Advances in nanomaterial-mediated CRISPR/Cas delivery: from lipid nanoparticles to vesicle-derived systems

Gene and genome editing therapies are increasingly connected with nanomaterials, which protect and transport fragile nucleic acids and CRISPR/Cas systems through biological barriers safely and accurately. This review discusses how different nanocarriers, including lipid-based, polymeric, inorganic, and vesicle-derived systems, can improve delivery efficiency, cell targeting, endosomal escape, and intracellular movement for gene and genome editing. It summarizes findings from early clinical and preclinical studies, comparing several carrier types such as ionizable lipid nanoparticles, polymeric nanoparticles, micelles, gold and silica nanostructures, and engineered extracellular vesicles. The review also explains how specific design factors, such as surface ligands, charge modification, PEGylation, and stimulus-responsive behaviors, influence biodistribution, and improve on-target efficiency while lowering immune responses and off-target effects. Ethical and regulatory concerns for in vivo editing are highlighted, along with current methods used to study nano–bio interactions. Among these carriers, ionizable lipid nanoparticles show the most advanced performance for delivering nucleic acids and CRISPR systems. However, new polymer-based and exosome-inspired carriers are progressing rapidly for repeated and targeted applications. Hybrid and responsive systems may also enable better spatial and temporal control of editing. Future research should focus on stronger in vivo potency testing, improved biocompatibility evaluation, and standardized manufacturing to ensure clinical safety and reliability.

1 Introduction

In genetic medicine, the integration of nanomaterials with gene and genome editing has enabled a major breakthrough, creating new possibilities for improving therapeutic strategies (Mostafavi and Zare, 2022). Over the past few decades, deeper understanding of the human genome has enabled the development of new methods to treat genetic disorders (Zhang W. et al., 2021). The use of nanomaterials has become a key innovation in this field, improving the precision, efficiency, and safety of gene therapy and genome editing (Yadav K. et al., 2023). Their unique physical and chemical properties make them suitable for developing more specific and targeted treatments (Liu L. et al., 2021), as they can interact directly with genetic material and influence how genes are controlled and expressed (Kim et al., 2018; Priyadarsini et al., 2018). Gene therapy, which involves adding, deleting, or modifying genes within human cells, offers hope for treating inherited diseases and certain cancers (Mirón-Barroso et al., 2021; Gong et al., 2023). However, its success depends on creating delivery systems that can transfer therapeutic genes safely and effectively to the target cells (Rizeq et al., 2019). Due to their adjustable size, surface charge, and high biocompatibility, nanomaterials have emerged as powerful carriers that help overcome these delivery challenges (Nocito et al., 2021).

Many types of nanomaterials have been explored to improve gene delivery, with each system providing distinct advantages (Dasari Shareena et al., 2018). Nanoparticles made from polymers or lipids can effectively encapsulate genetic material and protect it from enzymatic degradation (Nocito et al., 2021). Liposomes, composed of lipid bilayers resembling cellular membranes, facilitate cellular uptake by mimicking natural membrane structures (Jahangiri-Manesh et al., 2022). Engineered viral vectors also play a critical role in introducing therapeutic genes into target cells (Wei Hu et al., 2023). These nanomaterials not only safeguard the genetic cargo but also enhance intracellular transport, ensuring precise delivery to the intended site (Fernando et al., 2020). Meanwhile, CRISPR-based genome editing has revolutionized precision medicine (Gong et al., 2023). The CRISPR/Cas9 system allows for site-specific DNA modification with remarkable accuracy, offering strong potential for correcting genetic mutations and treating diseases at their molecular roots (Krasteva and Georgieva, 2022). However, the efficient delivery of CRISPR components to target cells remains a significant challenge (Shishparenok et al., 2023). Nanomaterials are essential in addressing this issue, as they can transport and protect CRISPR elements while guiding them to the appropriate genomic locations (Shi et al., 2014). Integrating nanomaterial-based carriers with CRISPR technologies enhances both editing precision and biosafety, advancing the practical use of these transformative tools (Hu et al., 2022).

The purpose of this research is to closely analyze the relationship between nanomaterials, gene therapy, and genome editing, focusing on current advances and potential future applications. The study mainly aims to explain how nanomaterials enhance the efficiency, precision, and safety of gene-based treatments and genome modification techniques. Various nanomaterial delivery systems, including nanoparticles, liposomes, and viral vectors, are examined to describe their functions in gene transfer, CRISPR-mediated genome editing, and RNA interference. Moreover, the review discusses the challenges associated with the use of nanomaterials in gene therapy, such as regulatory limitations and ethical concerns. It also identifies new developments and areas requiring further investigation. Overall, this study contributes to the expanding field of nanomaterial-based genetic medicine and supports a deeper understanding of their potential in treating genetic disorders.

2 Nanomaterial properties and functionalities

Engineered nanomaterials play a crucial role in enhancing the diagnosis and treatment of diseases by addressing the limitations of conventional drug delivery systems (Fernando et al., 2020). Nanotechnology introduces innovative strategies, including the targeting of specific cells and directing therapeutic molecules to designated organelles (Jahangiri-Manesh et al., 2022). These advances improve the precision of drug delivery and help resolve major challenges related to biodistribution and intracellular transport (Raya et al., 2022). In 2000, the U.S. National Science and Technology Council initiated the National Nanotechnology Initiative (NNI), which outlined strategic objectives and challenges to stimulate progress and promote the development of advanced nano-enabled technologies (Boverhof et al., 2015). Nanoparticles (NPs) demonstrate significant potential by enhancing cargo stability and solubility, facilitating membrane transport, and prolonging circulation time, which together contribute to safer and more effective therapies (Figure 1). Despite extensive research, a considerable gap remains between preclinical animal results and human clinical applications, primarily due to limited understanding of physiological and pathological differences (Judith and Vasudevan, 2022). Bridging this gap is essential for realizing the full clinical potential of nanomedicine (Kumar et al., 2021; Stater et al., 2021).

Figure 1

Figure 1. Roles of nanoparticles (NPs) in cancer. NPs can impede (green) or promote (red) cancer progression by directly affecting tumor cells and stroma, modulating immune responses, influencing metastasis, and altering angiogenesis.

The surface functionality of nanomaterials is crucial for targeted gene delivery in gene therapy, as it enables specific and controlled interactions with target cells (Anwar et al., 2020). In this process, nanoparticles are functionalized with ligands that selectively bind to receptors overexpressed on the surface of target cells, ensuring accurate cellular uptake and the precise delivery of therapeutic genes (Wang and Guo, 2016; Kreyling et al., 2010; Dufresne, 2017). When ligands possess high affinity for their receptors, nanomaterials can efficiently navigate the biological environment and reach the desired cells or tissues, hence minimizing off-target effects and enhancing therapeutic outcomes (Liu et al., 2022a; Ying et al., 2017). Surface modification thus allows researchers to customize ligands according to cellular characteristics, improving the specificity and efficiency of gene delivery systems (Manimekala et al., 2022). One of the greatest challenges in gene therapy is overcoming biological barriers such as the blood–brain barrier (BBB), which tightly regulates the movement of substances from the bloodstream into the brain (Kushwaha et al., 2022; Liu et al., 2022b; Chen et al., 2019). To address this, nanomaterials are engineered with surface ligands that interact with specific receptors or transport systems on BBB endothelial cells, facilitating receptor-mediated translocation into brain tissue (Chi and Liu, 2023; Xu and Liang, 2020; Ge et al., 2019). Careful selection and engineering of ligands remain essential, as they determine binding affinity, transport efficiency, and overall success in crossing complex biological barriers (Hao et al., 2022; Quader and Kataoka, 2017).

Surface functionality plays a crucial role in maintaining the stability of nanomaterials within complex biological environments, as it determines how these materials interact with biological components, particularly the immune system, which can strongly influence their circulation time and efficiency in gene delivery (Brüngel et al., 2023; Cheng et al., 2023). To improve their stability and biocompatibility, nanoparticles are often modified through surface coatings and charge adjustments (Vaidh et al., 2022). Coating nanoparticles with materials such as polymers or lipids forms a protective layer that prevents immune recognition and reduces opsonization, a process in which blood proteins adhere to the nanoparticle surface and mark them for clearance by phagocytic cells (Nain et al., 2022; Kladko et al., 2021). Minimizing opsonization extends the circulation time of nanomaterials, allowing them more opportunities to reach target cells and effectively deliver therapeutic genes (Liu MY. et al., 2021). Adjusting the surface charge also affects nanoparticle–biological interactions; neutral or slightly negative surfaces are preferred because they decrease nonspecific interactions and improve the “stealth” properties of the nanomaterials, thus enhancing their systemic stability and functional lifespan (Chen et al., 2019; Cheng et al., 2023; Liu et al., 2020). Furthermore, ligand selection is a key factor in achieving cell-specific targeting during gene delivery, as nanoparticles are engineered with surface ligands that can recognize and bind to overexpressed receptors on target cells (Rao et al., 2009; Liang T. et al., 2022). The process begins by characterizing the target cells and identifying receptors of interest, followed by selecting ligands with high affinity for those receptors (Yao et al., 2018; Jackman et al., 2021). This targeted recognition facilitates efficient cellular uptake, as the binding between the ligands and receptors triggers internalization mechanisms that enable the nanoparticles to enter the desired cells and deliver therapeutic genes (Huang et al., 2023; Buschmann et al., 2021). The specificity achieved through careful ligand selection helps minimize off-target effects by ensuring that nanoparticles interact only with intended cells, thereby reducing undesired biological responses and enhancing overall therapeutic efficacy (Bora et al., 2019; Yaqoob et al., 2020; Rawtani et al., 2019).

A biopolymer–Ni/Zn Np biocomposite (Prakasha et al., 2023) was developed using the exopolysaccharide of Rhodotorula mucilaginosa UANL-00IL, which facilitated the formation of polymorphic nanoparticles of 8–26 nm in size (Rahman et al., 2022). Unlike conventional nanocomposites with defined geometries, this material lacked a fixed structural shape, contributing to its unique physicochemical properties (Liang and Zhao, 2021). It demonstrated strong antimicrobial and antibiofilm activities against Staphylococcus aureus and Pseudomonas aeruginosa and in vivo assessments in male rats confirmed its biocompatibility, showing no toxicity at a dose of 24 mg/kg body weight (Rezk et al., 2020). Similarly, the AgNps/ZnONps nanocomposite was synthesized using an aqueous extract of the green alga Ulva fasciata, producing silver and zinc oxide nanoparticles of varying morphologies and dimensions (He et al., 2019). This composite exhibited significant antibacterial potential against Escherichia coli and Salmonella spp., particularly at elevated concentrations, and showed a synergistic effect when combined with antibiotics, suggesting its applicability in combination antimicrobial therapies (Raza et al., 2021; Vance et al., 2015). Another notable material, the tungsten carbide (Wc), silver (Ag), and copper (Cu) nanocomposite, was fabricated using commercially obtained nanoparticles of differing shapes and sizes (Chu et al., 2017). The mixture displayed potent antimicrobial efficiency against S. aureus and P. aeruginosa, confirming the collective role of its metallic components in enhancing bactericidal properties (Makabenta et al., 2021). Additionally, Ag/TiO₂ nanorods were synthesized using the horizontal vapor-phase growth (HVPG) technique (Fu et al., 2022). Their needle-like ends were capable of penetrating bacterial membranes, which substantially increased their antibacterial performance against S. aureus by causing direct structural disruption of the cell wall (Lin and Wang, 2015).

Another nanostructure, the Ag–Au/ZnO nanostructure, was synthesized with Justicia adhatoda plant extract (Karthikeyan et al., 2021). It demonstrated good antimicrobial activity against E. coli and S. aureus, showing the value of plant-based nanocomposites (Jones and Grainger, 2009). The α-BiO₂-ZnO nanostructure, made by chemical synthesis, produced a 1.5-cm inhibition zone against S. aureus at 1 mg/L. T-β-D-glu-ZnO Nps (trichoderma-β-D-glucan-zinc oxide nanoparticles) were spherical in shape and synthesized with a fungal mycelial water extract from Trichoderma harzianum (Tsoi et al., 2016). They inhibited the growth of S. aureus inside roundworms and also showed dose-dependent inhibitory effects on human pulmonary carcinoma A549 cells (Shin et al., 2023). ZnO/Fe₃O₄/rGO nanocomposites were rod-shaped and spherical (Besenhard et al., 2023). They showed stronger effects against E. coli than S. aureus, and adding reduced graphene oxide (rGO) increased antibacterial activity (Irshad et al., 2023). A second type of Ag/TiO₂ was made with an aqueous extract of Acacia nilotica (Zhao et al., 2023). This nanocomposite produced inhibition zones of different sizes against E. coli, MRSA, and P. aeruginosa (Teleanu et al., 2019). Its mechanism included lowering glutathione levels, which caused ROS production and lipid peroxidation (Eichhorn et al., 2022). Finally, ZEO-Ag Nps, ZEO-Cu Nps, and ZEO-Zn Nps were coatings made from copper-doped chitosan–gelatin (CSG) nanocomposites prepared by green synthesis (Yan et al., 2020). Their antibacterial activity depends on copper levels, and tests showed no harm to bone marrow stromal cell functions on Cu-doped coatings. This improved the biological performance of Ti-based materials (Bokov et al., 2021). Surface charge modification is an important factor in how nanomaterials work in targeted gene delivery (Sohaebuddin et al., 2010). The charge of nanoparticles influences how they interact with biological systems, including circulation time and uptake by cells (Besenhard et al., 2023). By changing the surface charge, scientists can improve their performance for delivering therapeutic genes (Lee et al., 2020). Neutral or slightly negative charges are usually preferred for targeted gene delivery (Mohsin et al., 2023). This helps reduce unwanted interactions with proteins or cells in the blood, giving nanomaterials more time to reach target cells (Mohsin et al., 2023; Xie et al., 2019). Surface charge also affects cellular uptake (Min et al., 2021). By lowering electrostatic repulsion between negatively charged cell membranes and the nanoparticles, positively or neutrally charged surfaces can improve uptake (Ashish and Singh, 2021). This is important for gene delivery because higher cellular uptake raises the chance of successful delivery of therapeutic material (Kuhlbusch et al., 2018). Biodegradation is another challenge in nanoparticle gene therapy since therapeutic nucleic acids can break down before they reach target cells (Kim et al., 2022), reducing their effectiveness (Kabir et al., 2019). To solve this, researchers use stable and biocompatible materials, such as lipid-based nanoparticles or polymer carriers (Sethuram et al., 2022). These materials protect the genetic cargo from enzymes, keeping it stable during circulation and helping deliver it to the right tissues (Figure 2).

Figure 2

Figure 2. A nanoparticle for gene therapy. Reprinted from Chen et al. (2016) with permission from Springer Nature.

3 Advances in gene-based therapies

Nanomaterials can encapsulate therapeutic genes, protecting them from degradation and facilitating their controlled release (Naskar and Kim, 2020). Additionally, surface modifications of nanomaterials allow for improved cellular uptake and intracellular trafficking of therapeutic genes (Jeong et al., 2022). This targeted and controlled delivery helps optimize the therapeutic effect while minimizing potential side effects (Nafees et al., 2013). Moreover, nanomaterials can overcome physiological barriers such as the blood–brain barrier, enabling the delivery of gene therapies to previously inaccessible tissues (Sharma et al., 2022). These advances in delivery technology open new possibilities for treating diseases with a genetic component that were previously challenging to effectively address (Alamri et al., 2020). One example of nanomaterial-enhanced gene therapy is the use of liposomal nanoparticles (Pokhrel et al., 2016). These lipid-based nanomaterials have demonstrated success in delivering therapeutic genes, especially for treating diseases like cancer (Sheikhzadeh et al., 2021). Liposomal nanoparticles can encapsulate genes, protect them during circulation, and release them specifically at the target site, maximizing the therapeutic effect while minimizing damage to healthy tissues (Kishore et al., 2023). Multifunctional nanoparticles play a crucial role in preclinical gene delivery studies, offering a versatile platform for enhancing the efficiency and specificity of gene targeting (Zhang Y. et al., 2021). One key strategy in nanoparticle functionalization involves PEGylation, which employs entities such as PEG-βCD and PEG-PEI (Stern et al., 2012). This approach contributes to enhanced stabilization, prevents unwanted protein absorption, and extends circulation time, thereby optimizing the overall performance of gene delivery systems (Han et al., 2022). Another significant aspect of multifunctional nanoparticles lies in their targeting capabilities (Yin et al., 2017). Utilizing constructs such as RGD-HA-PEI-PBLG, R-PEG20C, and transferrin-coated lipid, researchers have achieved improved gene target efficacy in vivo (Yang et al., 2015). These targeted delivery systems enable a more precise and efficient approach, enhancing the therapeutic impact of gene delivery (Wang et al., 2018). Stimulus-responsive nanoparticles further enhance gene delivery efficacy (Yang TC. et al., 2020). Through pH-sensitive, light-sensitive, and redox-sensitive designs, these nanoparticles respond dynamically to the microenvironment, ensuring optimal gene delivery under specific conditions (Pillai, 2014). This adaptability enhances the overall success of gene delivery systems in vivo (Rawtani et al., 2019). Cell-penetrating nanoparticles, such as p (DAHa-E/APIb), exhibit the ability to cross cell membranes efficiently, facilitating enhanced cellular uptake (Unnisa et al., 2023). This property is vital for ensuring the effective delivery of genetic material into target cells, thereby maximizing the therapeutic potential of gene-based therapies (Yang Y. et al., 2020). In the context of endosome escaping, nanoparticles like (Arg)7-FI-PNA have demonstrated the ability to cross cell membranes and improve endosomal escaping, overcoming a significant barrier in the gene delivery process (Pillai and Ceballos-Coronel, 2013). This capability is critical for ensuring that the genetic material reaches its intended destination within the cell (Wong et al., 2023). Nuclear localization is another key consideration, and nanoparticles such as PC/pDNA/NLS and VKRKKKP-R8 have been designed to facilitate it (Samarasinghe et al., 2012). This capability ensures that the delivered genetic material reaches the cell nucleus, where it can exert its therapeutic effects more effectively (Åström et al., 2015). Table 1 gives a detailed summary of gene-based therapies that use nanomaterials to treat different diseases. One clear example is the use of polymeric nanoparticles in delivering gene therapies for genetic disorders (Zheng et al., 2021). These nanoparticles, made of biocompatible and biodegradable materials, have been shown to deliver therapeutic genes effectively to target cells (Vimbela et al., 2017). In Duchenne muscular dystrophy (DMD), a serious genetic disease, researchers have used polymeric nanoparticles to carry the dystrophin gene. This method restored gene expression and reduced disease symptoms in preclinical studies (Yuan et al., 2023). Another important example is the use of viral vectors covered with nanomaterials to improve their safety and efficiency (Flores et al., 2012). Adeno-associated virus (AAV), which is often used in gene therapy, has been coated with nanomaterials to increase stability and improve targeting (Suri et al., 2007). This improved AAV has shown good results in treating inherited retinal diseases by delivering therapeutic genes to the retina, slowing the loss of vision (Zhi et al., 2022). Lipid-based nanocarriers are also important in developing cancer gene therapies (Kavanagh and Green, 2022). For example, liposomal nanoparticles have been used to deliver tumor-suppressing genes directly to cancer cells (Sahel et al., 2023). This targeted approach reduces harm to healthy tissues and increases therapeutic effects, highlighting the value of nanomaterials in precision medicine for cancer treatment (Demirer et al., 2021).

Table 1

Table 1. Examples of gene-based therapies utilizing nanomaterials for various diseases.

One of the main ways nanomaterials reduce off-target effects is by surface modifications that improve cellular specificity (Xi et al., 2022). When nanomaterial surfaces are functionalized with ligands or peptides, they can selectively bind to specific cell receptors, directing the nanoparticles to the intended target cells (Mohammadinejad et al., 2020). This targeted process lowers the chance of therapeutic genes affecting non-target cells, thereby reducing off-target effects (Mohamad et al., 2023). In addition, the size and physicochemical properties of nanomaterials help them avoid the immune system and reach the target cells more effectively (Liu et al., 2023). Encapsulating therapeutic genes within nanocarriers protects them from degradation and immune detection during circulation (Xin et al., 2022). This protection ensures that the therapeutic payload remains intact until it arrives at the target site, thus reducing the possibility of off-target interactions (Duan et al., 2021). Nanomaterials also provide controlled release systems, which allow a gradual and continuous delivery of therapeutic genes (He and Zhao, 2020). Such controlled release is important because it prevents sudden increases in gene expression that might otherwise cause off-target effects (Tariq et al., 2023). By controlling release kinetics, nanomaterials help maintain a therapeutic concentration at the target site while reducing exposure in non-target tissues (Hamid and Salee, 2022). In addition, nanomaterials can cross physiological barriers, such as the BBB, which improves their specificity for particular tissues (Yang et al., 2022). This property is very important in gene therapies for diseases of the central nervous system since off-target effects may cause serious problems (Zhang L. et al., 2022). The BBB is a natural barrier that limits the entry of foreign substances, including therapeutic agents, into the brain (Höijer et al., 2020). Although it is essential for keeping the central nervous system stable, the BBB also creates a major obstacle to delivering gene therapies to treat neurological diseases (Dodds, 2023). Nanomaterials can overcome this obstacle through several mechanisms (Hossain A. et al., 2021). One common method is to modify the surface of nanoparticles with ligands or peptides that can attach to receptors on BBB endothelial cells (Zhen et al., 2020). These modified nanoparticles are able to bind to the receptors, cross the BBB by transcytosis, and deliver therapeutic genes into the brain parenchyma (Foley et al., 2022). This strategy increases the specificity of gene delivery and reduces off-target effects in other tissues (Saritha et al., 2022). The small size of nanomaterials is very important for their ability to pass through the tight junctions of the BBB (Cho et al., 2019). Nanoparticles can use endocytosis and transcytosis pathways to cross the BBB effectively, which makes it possible to deliver therapeutic genes into the brain (Aziz et al., 2023). This feature is highly useful for treating neurodegenerative diseases and genetic disorders that affect the central nervous system (Campuzano and Pingarrón, 2023). In addition, nanocarriers help protect therapeutic genes while they circulate in the body (Huang et al., 2021). The encapsulation of genes inside nanomaterials protects them from enzymatic breakdown and immune system detection, keeping them safe and functional when they reach the target site in the brain (Shende and Trivedi, 2021). One of the main functions of liposomal nanoparticles is their capacity to enclose and protect therapeutic genes (Foley et al., 2022). The lipid bilayer of liposomes forms a safe environment around the gene cargo, preventing enzymatic breakdown and avoiding immune detection during circulation in the body (Chen et al., 2020). This protective role helps ensure the stable and efficient delivery of therapeutic genes to the tumor site (Yang et al., 2022). In addition, the surface properties of liposomal nanoparticles can be adjusted to improve their selectivity toward cancer cells (Sharma and Lew, 2022). By adding targeting ligands or antibodies, they can specifically recognize and attach to receptors that are highly expressed on the surface of cancer cells (Satta et al., 2023). This targeted strategy allows greater build-up of liposomal nanoparticles inside tumor tissue, which improves therapeutic impact while reducing harm to normal healthy cells (Lee et al., 2022). Liposomal nanoparticles can also provide the controlled release of therapeutic genes, which allows for longer exposure to cancer cells (Yan and Liang, 2022). This slow release is important because it helps keep an effective number of therapeutic genes in the tumor microenvironment, thus improving treatment results (Liang and Liang, 2021). In addition, liposomal nanoparticles are flexible and can deliver more than one therapeutic agent, such as genes together with chemotherapy drugs (Crowley et al., 2021). Using this combined strategy increases the success of cancer treatment by targeting different parts of tumor growth and development at the same time (Chin et al., 2019). Progress in gene-based therapies has also changed personalized medicine, making it possible to design treatments that match the genetic background of each patient (Zhang A. et al., 2022). A major benefit of gene-based therapies in personalized medicine is that they can address genetic disorders at their source (Tiwari et al., 2023). Instead of only managing symptoms, these treatments focus on the genetic mutations or problems that cause the disease (Li et al., 2022a). This approach provides more effective and lasting results, which fits with the main ideas of personalized medicine (Liang Y. et al., 2022). Gene therapies also make it possible to design treatments based on each person’s unique genetic profile (Abinaya and Viswanathan, 2021). By studying a patient’s genetic data, doctors can find specific markers or mutations that play a role in the development of disease (Gbian and Omri, 2021). Tailoring gene therapies to focus on these specific genetic factors can improve the effectiveness of treatment while reducing possible side effects, creating a more personalized type of patient care (Souri et al., 2022). In addition, progress in gene editing tools, especially CRISPR/Cas9, has introduced new opportunities for accurate genome changes (Lu et al., 2023). This progress makes it possible to fix harmful genetic mutations or add therapeutic genes directly into the patient’s genome (Rouatbi et al., 2019). Editing genes at the molecular level increases the value of personalized medicine because it addresses the exact genetic causes of diseases (Evans et al., 2019). The use of gene-based therapies is not limited to genetic disorders but also extends to cancer treatment (Moradi et al., 2022). In this case, targeted gene therapies for cancer can use the unique genetic features of tumor cells, giving a specific approach that avoids unnecessary harm to healthy tissues (Wu Z. et al., 2022). Such targeted accuracy is a main feature of personalized medicine, where treatment plans are created based on the genetic and molecular profile of each individual (Dinari et al., 2018).

3.1 Precision genome editing techniques

Nanomaterials have an important role in improving precision genome editing because they act as carriers for different gene-editing tools (QIAO et al., 2021). They are usually designed as nanoparticles and display special features such as large surface area, good biocompatibility, and the possibility of surface modification (Chenouard et al., 2023). These properties make nanomaterials, such as gold nanoparticles, suitable for delivering CRISPR/Cas9 components with accuracy, which supports efficient and targeted changes in the genome (Khan, 2021). Their function in protecting and transporting genetic material also increases the success of precision genome editing methods (Sun et al., 2023). Many types of nanomaterials are useful in this field, including liposomes, polymers, and inorganic nanoparticles such as gold and silica (Khan, 2021). Liposomes—made of lipid bilayers—can encapsulate and transport genetic material, while biodegradable polymers are helpful for the controlled release of editing tools (Sun et al., 2023). Inorganic nanoparticles like gold and silica give stability and controlled release because of their tunable surface chemistry and porous structure (Ojha et al., 2023). Nanomaterials thus improve the outcomes of genome editing applications (Tay and Melosh, 2019; Wang et al., 2025). CRISPR technology, which uses RNA-guided Cas proteins, has changed the field of precision genome editing (Ryu et al., 2020). It gives researchers the ability to edit genes precisely, either to correct mutations or add specific changes (Shen Y. et al., 2018). Despite being simple, cost-effective, and versatile, CRISPR still faces challenges such as off-target effects and ethical issues, which have pushed researchers to improve it further (Li et al., 2017). Although CRISPR technology is the leading tool, other genome editing methods like zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) are also available (Wei et al., 2022). Both ZFNs and TALENs are protein-based systems that create double-strand breaks for editing (Yang et al., 2023). However, compared to CRISPR, they are usually more complex and less flexible in their design (Li, 2019). The ease of use and versatility of CRISPR make it the most common choice for precise genome editing, though scientists continue to work on making it more accurate and on reducing unwanted effects (Zhai et al., 2022).

Nanomaterials play an important role in delivering CRISPR components by encapsulating and protecting genetic material until it reaches target cells (Alsulami, 2021). For example, lipid-based nanocarriers form stable complexes with CRISPR, protecting them from degradation and helping cellular uptake (Zia et al., 2021). The surface properties of nanomaterials can also be designed to improve targeting, reduce off-target effects, and increase precision in genome editing (Peng et al., 2020). Researchers are still improving nanomaterials to make delivery safer and more effective (Gao et al., 2021). By modifying surfaces and adding functional groups, they aim to lower toxicity and improve compatibility with cells (Roma-rodrigues et al., 2020). In addition, stimulus-responsive nanomaterials allow the controlled release of CRISPR at specific places inside cells; this supports the development of safer nanomaterial-based delivery systems and helps address current challenges (Dannhäuser et al., 2022).

CRISPR technology solves many of the challenges of older gene-editing tools such as zinc finger nucleases and TALENs by offering a simpler and more flexible approach (Yadav MR. et al., 2023). The easy design of guide RNAs for chosen DNA sequences improves accuracy, making CRISPR a cost-effective and scalable method. This is why researchers and clinicians prefer it for creating precise and efficient genome modifications (Sharshar et al., 2020). In addition to correcting genetic diseases, precision genome editing can be used to create genetically modified organisms in agriculture, produce disease-resistant crops, and design microbes for better biofuel production (Niazian et al., 2021). It also helps scientists study gene function and regulation, which supports new knowledge about biological processes (Edagwa et al., 2017). Its possible uses in cancer immunotherapy and treatment of infectious diseases show how powerful and wide-ranging this technology is for both science and industry (Liu et al., 2018). However, avoiding off-target effects with CRISPR remains a key concern (Shen et al., 2021). Current research is working to improve CRISPR accuracy through better guide RNAs and modified Cas proteins (Ma et al., 2017). In addition, bioinformatics tools are used to predict possible off-target sites, which helps scientists design improved guide RNA sequences (Shen et al., 2022). Innovative CRISPR variants that show reduced off-target effects are being developed, and this supports the continued progress and reliability of CRISPR technology (Pezoa et al., 2022). A detailed understanding of the factors that influence off-target effects enables researchers to design strategies that can reduce these problems (feng et al., 2023). This commitment helps overcome one of the major limitations of CRISPR and improves its reliability (Fiaz et al., 2021). At the same time, nanomaterial-based delivery systems are advancing, and researchers are working to make them safer and more effective (Ladu, 2021). Surface modifications and functionalizations are especially important in lowering cytotoxicity and improving biocompatibility, thus making these nanocarriers more dependable for precise genome editing (Mushtaq et al., 2019). Another important step is the creation of stimulus-responsive nanomaterials which allow CRISPR components to be released in a controlled way inside specific target cells (Doroftei et al., 2021). Such precision makes genome editing techniques more accurate and reduces the risks linked to off-target effects (Corte et al., 2019). In genome editing approaches, CRISPR remains dominant because it is simple, flexible, and cost-effective (Ferdous et al., 2022). Its programmable nature allows scientists to adapt it to different genetic modifications, making it a suitable choice for many applications (Khan, 2019). Other tools, such as ZFNs and TALENs, are available, but their complex design and reduced flexibility make them less widely used than CRISPR (Katayama et al., 2016). The applications of precision genome editing go far beyond fixing genetic diseases (Popova et al., 2023). This technology also has great value in agriculture, where genetically modified organisms are created to increase crop yields and increase plant resistance to disease (Calos, 2017). It is also used for engineering microbes that can improve biofuel production, thus supporting sustainable energy solutions (Haque et al., 2021). In addition to these industrial uses, precision genome editing helps scientists study gene function and regulation, which leads to progress in many biological processes (van de Wiel et al., 2017). The medical use of genome editing is also clear in cancer immunotherapy and in treating infectious diseases (Feng et al., 2018). Because it can change genes very accurately, it gives researchers new ways to develop targeted therapies that may change how diseases are treated (Zych et al., 2018). As studies continue, the wide use and strong impact of genome editing become more obvious, influencing the future of both medicine and biotechnology (Maity et al., 2023). A strong connection between nanomaterials and genome editing has also opened new opportunities (Agapito-Tenfen, 2016). Nanomaterials, with their special properties, make it possible to deliver gene-editing tools more effectively and with higher precision, solving problems in methods such as CRISPR (Su et al., 2016). As new delivery systems using nanomaterials improve and CRISPR technology itself becomes more advanced, genome editing will likely transform medicine, agriculture, and many other industries (Lamboro et al., 2021).

3.2 Nanomaterial-based delivery systems

Nanomaterial-based delivery systems play an important role in enhancing the targeted delivery of therapeutic agents by using the special properties of nanoparticles, liposomes, and viral vectors. Figure 3 shows the mechanisms through which nanoparticle-related effects influence cancer (Gogolev et al., 2021). Because of their very small size, nanoparticles can pass through biological barriers, reach certain cells or tissues, and facilitate accurate drug delivery (Ansari et al., 2020). The lipid bilayer structure of liposomes is similar to cell membranes, which helps in cellular uptake and provides the controlled release of therapeutic agents (Zhou et al., 2022). Viral vectors, which are safely modified from viruses, act as highly effective carriers that deliver genetic material to target cells with strong accuracy. Figure 4 presents the use of different analytical methods for studying nano–bio interactions at various levels.

Figure 3

Figure 3. Mechanisms by which nanoparticle-induced ancillary effects impact cancer. (a) Ferumoxytol iron oxide nanoparticles modulate macrophage polarization in tumors, promoting M1 polarization and inducing cancer cell death through extracellular reactive oxygen species (ROS). (b) Uptake of ferumoxytol by iron-retaining leukemia cells activates the ferroptotic pathway, leading to cell death. (c) Titanium dioxide nanoparticles alter the permissiveness of endothelial cells in the microvasculature, thereby facilitating metastasis. (d) Carbon dots (C-dots) accumulate iron in vivo, triggering ferroptotic cell death in cancer cells. FPN, ferroportin. Reprinted from Stater et al. (2021) with permission from Springer Nature.

Figure 4

Figure 4. Analytical methods used to study nano–bio interactions at different levels. Diagram shows (a) surface plasmon resonance (SPR) as an indirect method for analyzing the biocorona; (b) ATR-FTIR spectra that monitor glycine adsorption on TiO₂ nanoparticles over time; (c) QCM-D spectra that present frequency changes during fibrinogen adsorption on surfaces with different topographies; (d) fluorescence correlation spectroscopy (FCS) as an indirect method to study nanomaterial–protein interactions; (e) synchrotron radiation circular dichroism (SR-CD) spectra of HSA in PBS before and after contact with nanosheets (NSs); (f) schematic view of the internalization pathway of cubic PEGylated nanoparticles; (g) transmission electron microscopy (TEM) images of cells and Ag nanoparticles; (h) effect of protein corona on cell adhesion and survival; (i) soft X-ray transmission microscopy (STXM) dual-energy contrast images displaying macrophage uptake of Gd@C₈₂(OH)₂₂ in vivo; (j) X-ray absorption near-edge structure (XANES) analysis of silver chemical forms during uptake and clearance in cells; (k) flow cytometry used to detect exposed TGF-β1 in the protein corona; (l) multi-omics analysis for toxicity evaluation in risk assessment; (m–o) quantification of in vivo nanomaterial distribution using inductively coupled plasma mass spectrometry (ICP-MS), single-photon emission computed tomography/computed tomography (SPECT/CT), and magnetic resonance imaging (MRI); (p) laser ablation–ICP-MS gold imaging showing PEG-GNP movement and intra-organ distribution in the liver; (q, r) synchrotron radiation X-ray fluorescence (SR-XRF) microscopy and XANES mapping of molybdenum in the liver together with the characterization of in vivo biotransformation of MoS₂. Reprinted from Liu et al. (2022a) with permission from Elsevier.

Nanoparticles offer several advantages in drug delivery, making them indispensable in the field. Their small size enables efficient cellular uptake, delivering therapeutic agents directly into target cells (Joun et al., 2022). The high surface-area-to-volume ratio allows for functionalization, tailoring surface properties for specific applications (Rong et al., 2020). Additionally, tunable release kinetics ensure controlled and sustained drug release, thus optimizing therapeutic efficacy (Ibrahim et al., 2022). Biocompatible and biodegradable nanoparticle formulations enhance their safety profile, making them ideal carriers for various therapeutic compounds (Turcheniuk and Mochalin, 2017a). Table 2 presents an overview of nanomaterial-based delivery systems, showcasing various advances and applications in the field.

Table 2

Table 2. Nanomaterial-based delivery systems.

3.3 Chitosan

Polymeric nanoparticles show high biocompatibility and adjustable properties, giving them a long residence time in ocular tissues (Lin et al., 2022). They can carry both hydrophilic and hydrophobic drugs, which makes them useful for different therapeutic agents (Liu J. et al., 2021). Their ability to provide sustained release allows for longer therapeutic effects (Banerjee et al., 2019). In addition, they can use both passive and active targeting, improving their value in treating conditions such as glaucoma, age-related macular degeneration, and diabetic retinopathy (Dubey et al., 2023). However, despite these benefits, polymeric nanoparticles also face some limitations (Velluto et al., 2021). They may aggregate, and their loading capacity for hydrophobic drugs is limited (Jain, 2012). These problems can reduce drug delivery efficiency and must be carefully considered during formulation (Miao et al., 2014). Even so, current research is trying to solve these problems and expand the use of polymeric nanoparticles in ocular drug delivery (Li et al., 2022b).

Liposomes, although unstable and costly to produce, are biocompatible and allow good control over drug release (Štimac et al., 2019). They are better suited for hydrophobic drugs and can provide sustained release, making them useful for conditions such as dry eye syndrome and fungal keratitis (Soroush et al., 2016). Passive targeting is possible, but their stability problems encourage more studies to improve their design and manufacturing (Singh et al., 2013). Chitosan-based implants provide sustained release, good biocompatibility, and biodegradability, which make them effective for long-term treatment (Yu et al., 2016a). The main disadvantage is that they require surgical implantation (Dash et al., 2021). Nevertheless, because of their extended drug release and compatibility, they are promising for treating ocular diseases such as glaucoma and retinoblastoma (Arjama et al., 2018). Chitosan-coated prodrugs improve drug solubility and stability, and they can deliver drugs directly to ocular tissues (Yu et al., 2016a). Their complex design and synthesis are challenges, but their controlled release and potential for active targeting make them valuable for ocular diseases (Aseri et al., 2015). These prodrugs show strong potential in drug delivery, especially for conditions that need better solubility, stability, and targeted release (Santana et al., 2022).

3.4 Alginate

In oral drug delivery, alginate has an important role as it is used in tablets and capsules for delivering proteins, peptides, and nucleotides (Ashique et al., 2023). The release system is mainly pH-sensitive, which makes it possible to deliver drugs to selected parts of the gastrointestinal tract (Hastings et al., 2015). Alginate is often chosen because it is biocompatible and non-toxic, making it a good material for controlled and sustained drug release (Madl et al., 2012). However, alginate can also face the problem of gel erosion, which may require extra solutions such as enteric coatings (Pan et al., 2021). These coatings stop the drug from being released too early and help ensure proper delivery (Gupta, 2020). In practice, alginate is applied in the form of tablets and capsules for proteins, peptides, and nucleotides (Khan et al., 2023). The release system works in a pH-sensitive way, allowing targeted release in certain regions of the gastrointestinal tract (Yadav P. et al., 2023). Because it is safe and non-toxic, alginate is a suitable material for achieving long-term and steady drug release (Lan et al., 2020). Despite these benefits, the problem of gel erosion when using alginate may require the use of enteric coatings (Tang et al., 2010). These help prevent early drug release and support proper drug delivery (Yu et al., 2016b). In oral drug delivery, alginate is used in making tablets and capsules for delivering proteins, peptides, and nucleotides (Kotb et al., 2023). The main release method is pH-sensitive, which ensures drug delivery to exact areas of the gastrointestinal tract (Dai et al., 2021). Alginate is chosen because it is biocompatible and non-toxic, which makes it suitable for controlled drug release (Khan et al., 2023). However, the risk of gel erosion linked with alginate may make enteric coatings necessary (Manikantan et al., 2023). These coatings work as a protective layer, stopping early drug release and helping the drug delivery system to remain effective (Chen et al., 2017). In oral drug delivery, alginate is applied in preparing tablets and capsules that carry proteins, peptides, and nucleotides (Xavier Mendes et al., 2021). The use of a pH-sensitive system guarantees targeted drug delivery to the right parts of the gastrointestinal tract (Shafee et al., 2021). Because alginate is safe and non-toxic, it is a strong option for continuous drug release (Cai and Xu, 2011). Nevertheless, the issue of gel erosion in alginate use raises the need for extra methods, such as adding enteric coatings (Pillai et al., 2018). These coatings act as protection, avoiding early release of drugs and keeping the drug delivery process effective (Turcheniuk and Mochalin, 2017b). In oral drug delivery, alginate plays an important role in making tablets and capsules for proteins, peptides, and nucleotides (Kausar, 2021). The main idea of its release mechanism is based on pH sensitivity, which allows drugs to be released carefully and only in certain parts of the gastrointestinal tract (Lin and Mao, 2011). Alginate has many useful properties, such as being biocompatible and non-toxic, which make it suitable for controlled drug release (Barua et al., 2020). However, one problem with alginate is gel erosion, and this requires a practical solution, such as using enteric coatings (Kang et al., 2017). These coatings act as a barrier to protect the drug, lowering the chance of early release and keeping the accuracy and effectiveness of the delivery process (Barhoum et al., 2022). In oral drug delivery, alginate is an important material used in tablets and capsules designed to release proteins, peptides, and nucleotides (Lupu et al., 2020). The key principle of its release system depends on pH sensitivity, which helps the drug reach specific areas of the gastrointestinal tract (Szczeszak et al., 2015). Because alginate is biocompatible and non-toxic, it is a good choice for supporting controlled and sustained drug release (Vélez et al., 2017). Still, the risk of gel erosion when using alginate makes it necessary to find other solutions, such as enteric coatings (Ohyanagi et al., 2011). These coatings work as a protective shield, reducing the chance of drugs releasing too early and making sure drug delivery remains precise and effective (Hossain CM. et al., 2021).

3.5 Liposomes

Liposomes and alginate are two distinct systems in the field of drug delivery, each defined by specific characteristics and applications (Kapoor et al., 2018). Structurally, liposomes are spherical vesicles that consist of a phospholipid bilayer membrane, while alginate is a linear polysaccharide composed of β-D-mannuronic acid and α-L-guluronic acid residues (Giordani, 2020). The architecture of liposomes is more complex than alginate (McDonald et al., 2015). This structural complexity, however, allows liposomes to provide greater flexibility in modifying their composition for targeted drug delivery (Opanasopit et al., 2008). In contrast, alginate is a natural and biodegradable polymer, which highlights its environmentally friendly properties (Amreddy et al., 2017). With respect to charge, liposomes may be neutral, cationic, or anionic, making it possible to adjust their surface charges for specific cellular interactions (Zhou et al., 2018). For example, cationic liposomes can improve cellular uptake through electrostatic attraction (Khaliq et al., 2023). Alginate, although typically non-ionic, can be engineered into ionic forms, demonstrating its adaptability in charge modification (Han et al., 2021). Among the major benefits of liposomes is their capacity to be functionalized with ligands, enabling targeted delivery to particular cells and offering additional strategies for selective drug administration through ligand conjugation (Mashima and Takada, 2022). Alginate, unlike liposomes, can reach particular tissues through passive targeting mechanisms (Majidi et al., 2016). Liposomes may show instability under physiological conditions, which creates the need for structural modifications to enhance their stability (Wang et al., 2015). In contrast, alginate is normally stable in physiological environments, making it a stronger option in this aspect (Luly et al., 2020). Both liposomes and alginate are biocompatible. Liposomes are also mucoadhesive, which makes them appropriate for wound healing and for drug delivery in the gastrointestinal tract (Khaliq et al., 2023). The drug loading ability of these carriers shows different features (Xu et al., 2022). Liposomes can encapsulate both hydrophilic and hydrophobic drugs, and they provide higher efficiency for a wide range of molecules (Tian et al., 2013). Furthermore, liposomes can be engineered for sustained or controlled release (Pissuwan et al., 2011). Alginate, mostly used for hydrophilic drugs, can form hydrogels that allow localized drug delivery (Zhu et al., 2022). Applications of liposomes include drug delivery, hydrophobic drug encapsulation, gene therapy, and cosmetic products (Khaliq et al., 2023). Alginate, on the other hand, is widely applied in cell encapsulation, tissue engineering, and controlled release systems (Shen L. et al., 2018). Toxicity must also be considered in comparison. Liposomes are mostly well-tolerated but may cause dose-dependent toxicity (Shafee et al., 2021). Alginate is generally non-toxic, although it may trigger some immune response (Erel-Akbaba et al., 2021). It is often described as less toxic than certain liposome formulations, strengthening its safety profile (Lai et al., 2017). Both materials are important for drug delivery: Liposomes offer versatility and targeting potential, while alginate provides cost-effectiveness and mechanical strength for applications such as tissue engineering, cell encapsulation, and pharmaceutical systems (Lin et al., 2015). Cost is another critical factor (Casper et al., 2023). Liposomes are usually more costly to produce, while alginate is cheaper and therefore more suitable for large-scale pharmaceutical production (Davis, 2009). This cost advantage makes alginate an appealing alternative in broader drug delivery applications (Luly et al., 2020). The targeting abilities of liposomes extend beyond charge modification and include functionalization with ligands for specific cellular interactions (Deng et al., 2019). This property enables liposomes to provide more strategies for targeted drug delivery through ligand conjugation (Khanna et al., 2023). In comparison, alginate, although less versatile in this aspect, compensates with its ability to achieve tissue-specific delivery via passive targeting mechanisms (Lima et al., 2019). These differences demonstrate the unique strengths of each material in overcoming challenges in drug delivery (Naik et al., 2022). The application context further highlights the complementary roles of liposomes and alginate (Wang et al., 2021). Liposomes are particularly effective in drug delivery, especially for encapsulating hydrophobic drugs, in gene therapy, and in cosmetic formulations (Xu et al., 2021). Alginate, however, is mainly applied in cell encapsulation, tissue engineering, and controlled drug release (Shen Y. et al., 2018). The wide range of applications illustrates the distinct features each material contributes, confirming their importance in the biomedical field (Ghormade et al., 2011).

With regard to toxicity, liposomes are generally well tolerated, although dose-dependent toxicities can occur (Koupaei et al., 2019). By contrast, alginate is widely recognized as non-toxic, although it may trigger immune responses in certain cases (Kozielski et al., 2014). Its favorable safety profile, being less toxic than some liposome formulations, emphasizes its suitability for biomedical use where minimizing adverse effects is critical (Morán et al., 2018). Overall, liposomes and alginate present specific advantages and characteristics in the field of drug delivery (Wang et al., 2017). Liposomes are notable for their versatility, ability to manipulate charge, and potential for ligand-mediated targeting (Liang et al., 2023). Alginate is distinguished by its natural, biodegradable qualities, cost-effectiveness, physiological stability, and its applications in tissue engineering and cell encapsulation (González-Reyna et al., 2023). The combination of these advantages positions both liposomes and alginate as valuable elements in advancing drug delivery systems, with their individual attributes addressing different therapeutic and biomedical requirements (Wang et al., 2019).

4 Applications in CRISPR-based genome editing

Applications of CRISPR-based genome editing involve a wide variety of nanomaterials, each used through specific delivery methods to improve the precision and efficiency of gene modification (Imlimthan et al., 2020). Lipid nanoparticles, applied via encapsulation, have shown enhanced delivery performance and reduced off-target activity in human T cells (Zhao et al., 2020). In particular, editing the PD-1 gene, an immune checkpoint, produced stronger T cell responses against cancer cells, thus demonstrating significant potential for progress in cancer immunotherapy (Sami et al., 2012). Mesoporous silica nanoparticles, when conjugated with Cas9, provided greater stability and protection of the Cas9 enzyme (Qamar et al., 2021). This method, when applied to rice plants, successfully targeted the OsSWEET11 gene (a sugar transporter), producing increased drought resistance (Ihnatsyeu-Kachan et al., 2017). Carbon nanotubes, functionalized with guide RNA, displayed improved cellular uptake and efficient targeted delivery in mice (Liu et al., 2022a). By editing the HPRT1 (hypoxanthine–guanine phosphoribosyltransferase 1) gene, this approach corrected a genetic defect, serving as a model for therapeutic applications (Chen et al., 2017). Polymer nanoparticles carrying CRISPR components allowed controlled release and sustained delivery in bacteria (Xavier Mendes et al., 2021). Targeting multidrug resistance genes led to greater sensitivity to antibiotics, offering a potential strategy for overcoming antibiotic resistance (Shafee et al., 2021). Exosomes engineered for CRISPR delivery functioned as natural carriers with enhanced biocompatibility and selective targeting (Cai and Xu, 2011). In human stem cells, this approach was applied to the β-globin gene (hemoglobin subunit), indicating a possible treatment for sickle cell disease (Pillai et al., 2018). Magnetic nanoparticles manipulated by an external magnetic field have enabled spatially directed delivery to specific regions of zebrafish embryos. Editing the EGFP gene (fluorescent protein) permitted precise modifications in defined tissues, highlighting opportunities for spatially controlled genome editing (Priyadarsini et al., 2018). Gold nanoparticles, integrated with photothermal ablation and CRISPR, demonstrated a synergistic strategy against cancer cells. In human cancer cells, targeting the EGFR (epidermal growth factor receptor) gene enabled both precise editing and cancer cell destruction—an innovative method reported by Mirón-Barroso et al. (2021). Table 3 provides a summary of successful genome editing applications using nanomaterials.

Table 3

Table 3. Examples of successful genome editing applications using nanomaterials.

5 RNA interference

Nanomaterials act as important facilitators in RNA interference (RNAi) by creating a stable and protective environment for RNA molecules, such as small interfering RNA (siRNA) (Ferdous et al., 2022). Their unique features, including small size and specific surface characteristics, allow efficient encapsulation and delivery of siRNA to target cells (Su et al., 2016). This encapsulation protects the fragile RNA molecules from enzymatic degradation, increases their stability during transport, and improves their bioavailability once they reach the target cells (Katayama et al., 2016). The precision of nanomaterial-mediated RNAi is based on the ability to specifically regulate disease-associated genes, increasing the effectiveness of therapeutic interventions (Popova et al., 2023). Nanomaterials also influence the pharmacokinetics of RNA interference by improving the circulation and bioavailability of RNA molecules (Calos, 2017). Their small size supports longer circulation in the bloodstream and prevents rapid clearance by the immune system (Zhou et al., 2022). Furthermore, nanomaterials can be designed for enhanced permeability and retention (EPR), which enables selective accumulation at disease sites (van de Wiel et al., 2017). This accumulation raises the concentration of RNA molecules at the target, thus increasing therapeutic efficiency (Feng et al., 2018). By modulating the pharmacokinetics of RNA interference, nanomaterials improve precision and the overall success of therapies (Maity et al., 2023). Nanomaterials also offer a multifaceted approach to targeted delivery in RNA interference (Zych et al., 2018). Through surface modifications and functionalizations, they can be engineered to recognize specific markers on target cells, leading to selective binding and internalization (Agapito-Tenfen, 2016). This targeted delivery ensures the controlled release of RNA molecules at the intended sites and reduces off-target effects (Su et al., 2016). Such precision enhances therapeutic outcomes by concentrating the therapeutic load at the disease site, lowering systemic exposure and decreasing possible side effects (Lamboro et al., 2021). In addition, nanomaterials help overcome barriers linked to RNA interference (Gogolev et al., 2021). RNA delivery often faces problems such as enzymatic degradation, recognition by the immune system, and limited cellular uptake (Ansari et al., 2020). Nanomaterials address these challenges by offering protective encapsulation, shielding RNA from degradation, avoiding immune recognition, and improving cellular uptake (Zhou et al., 2022). These improvements strengthen the precision and effectiveness of therapeutic strategies by ensuring the successful delivery of RNA molecules to target cells while overcoming biological barriers (Singh et al., 2013). Different nanomaterials have been developed to support RNA interference, each enhancing the accuracy and efficiency of therapeutic methods (Agapito-Tenfen, 2016). Lipid-based, polymeric, and inorganic nanoparticles are notable examples (Padayachee and Singh, 2020). Lipid-based nanoparticles provide a lipid bilayer resembling cell membranes, which increases cellular uptake (Falzarano et al., 2015). Polymeric nanoparticles allow the modification of surface properties, making targeted delivery possible (Turcheniuk and Mochalin, 2017a). Inorganic nanoparticles, including gold nanoparticles, show distinctive physicochemical properties that support effective RNA delivery (Ding et al., 2022). The choice of nanomaterial depends on the specific therapeutic aim and the type of RNA molecules delivered. These nanomaterials illustrate the range of strategies used to increase precision in RNA interference and optimize therapeutic performance (Ghormade et al., 2011). Table 4 provides a detailed overview of nanomaterials and their role in RNAi. The wide variety of nanomaterials tested for RNA interference highlights the need to balance benefits and potential limitations for each type (Koupaei et al., 2019). Knowledge of the unique features and restrictions of each nanomaterial is essential for their best use in RNA interference applications (Kozielski et al., 2014). Lipid nanoparticles, which use a complexation loading method, are applied to different cell types (Rong et al., 2020). They show high delivery efficiency with controlled release and stable gene silencing (Mohamad et al., 2023). Their biodegradable structure and flexibility are advantages, but possible off-target effects and cost issues remain concerns (Wang et al., 2017). Carbon nanotubes, loaded through functionalization, are able to target cells that are otherwise hard to reach (Wang et al., 2017). Although their delivery efficiency varies and is still under study, they have strong potential for selective delivery (Liang et al., 2023). However, safety risks and aggregation remain major drawbacks (Song et al., 2023). Metal nanoparticles, applied through conjugation, can target a variety of cell types (González-Reyna et al., 2023). Their delivery efficiency is moderate and requires improvement, with gene silencing effects also at a moderate level (Wang et al., 2019). Their strengths are biocompatibility and adjustable properties, but disadvantages include toxicity and non-specific interactions (Imlimthan et al., 2020). Dendrimers, commonly loaded through encapsulation, are utilized for delivery across different cell types, showing delivery efficiency and gene silencing efficiency that depend on their size (Zhao et al., 2020). These nanoparticles are biocompatible and adaptable; however, potential toxicity and the difficulty of achieving controlled release remain important concerns (Sami et al., 2012). Exosomes, functioning as natural carriers, demonstrate strong delivery efficiency with inherent targeting across multiple cell types (Morán et al., 2018). Their advantages include biocompatibility and low immunogenicity, although their restricted loading capacity and variability between batches present challenges (Zhang L. et al., 2022). Inorganic nanoparticles, such as gold nanoparticles, are generally loaded through conjugation and are directed toward particular cell types (Khaliq et al., 2023). Their delivery efficiency continues to vary and remains the subject of ongoing research, as does their variable gene silencing efficiency (Dodds, 2023). These nanoparticles offer unique properties, including imaging functions, and display potential for controlled release (Lin et al., 2022). Nevertheless, safety issues and the incomplete understanding of their mechanisms represent significant disadvantages (Liu J. et al., 2021). Figure 5 illustrates the immunomodulatory effects achieved through nanoparticle mediation and how these contribute to antigen tolerance.

Table 4

Table 4. Examination of nanomaterials in facilitating RNA interference.

Figure 5

Figure 5. Immunomodulatory effects induced by nanoparticles (NPs) support the establishment of antigen tolerance. In the upper section, the foreign antigen is internalized by antigen-presenting cells (APCs) together with an external pro-inflammatory activation signal. This co-stimulation contributes to the priming of naive T cells, resulting in the activation and/or proliferation of mature antigen-reactive T cells. This occurs through enhanced presentation of antigen fragments via major histocompatibility complexes (MHCs), the expression of juxtacrine co-stimulatory receptors, and the release of pro-inflammatory cytokines. In contrast, the lower section illustrates the co-uptake of antigen with poly (lactic-co-glycolic acid) (PLGA) nanomaterial. The degradation of PLGA into acidic monomers leads to the inhibition of TAK1, mediated by lactic acid, which reduces NF-κB phosphorylation and prevents nuclear translocation of pNF-κB. Consequently, the responsiveness to pro-inflammatory signals acting through TAK1 is weakened. When sufficient co-stimulation is absent, the presentation of antigen fragments to T cells does not activate naive cells and instead promotes the senescence or apoptosis of mature antigen-reactive T cells. The figure also includes essential labels such as IFN-γ (interferon-γ), IL (interleukin), MHC (major histocompatibility complex), TCR (T-cell receptor), and TNF (tumor necrosis factor). Reprinted from Stater et al. (2021) with permission of Elsevier.

6 Clinical development of nanoparticles for gene delivery

Multifunctional nanoparticles have received considerable attention in preclinical studies related to gene delivery (Madl et al., 2012). The area of gene therapy, especially in the prevention and treatment of genetic diseases, has experienced a rapid increase in clinical trials worldwide (Pan et al., 2021). However, despite such progress, none of the gene therapeutics based on nanoparticles have yet gained approval from the FDA. This absence of approval shows the continuing difficulties and safety concerns linked to gene therapy (Gupta, 2020). A milestone in the clinical progress of gene therapy was achieved when Anderson et al. carried out the first human clinical trial, where the adenosine deaminase gene was systemically delivered to a 4-year-old girl suffering from severe combined immunodeficiency disease. The initial success of this case marked the beginning of a global expansion of research in gene therapy (Khan et al., 2023). Another important clinical trial was on severe combined immunodeficiency-X1. This inherited disorder, linked to the X chromosome, was treated using complementary DNA in a retrovirus-derived vector with ex vivo infection of CD34⁺ cells (Yadav P. et al., 2023). The 10-month follow-up showed encouraging outcomes, with T, B, and NK cell counts and their functions similar to those of age-matched healthy controls. Nevertheless, safety issues appeared later. After 3 years, the two youngest boys in the trial developed leukemia; this was mainly due to retrovirus vector integration near a proto-oncogene, which caused deregulated premalignant cell growth (Lan et al., 2020). This incident highlighted the safety risks of viral vectors and placed serious limits on the wide use of gene therapy (Tang et al., 2010). Despite these challenges, researchers continue to examine safer and more efficient strategies for gene delivery through multifunctional nanoparticles in preclinical studies, showing the active and developing character of this field (Yu et al., 2016b).

Nanoparticle-based gene therapy is currently under clinical evaluation for a wide range of diseases, using different delivery systems. One such approach is the use of polyethylenimine (PEI)-based nanoparticles. BC-819/PEI, developed by BioCanCell, is being studied for bladder cancer (BC) through local administration and is in phase 2 with active status (NCT00595088). In addition, BC-819, also from BioCanCell, has finished phase 1/2 clinical trials for ovarian cancer (OC) with intraperitoneal (IP) administration (NCT00826150). Another product, DTA-H19 by BioCanCell, has completed phase 1/2 trials for pancreatic neoplasms (PN) through local administration (NCT00711997).

Lipid-based nanoparticles represent another important system. TKM-080301, from Tekmira Pharmaceuticals Corporation, has completed phase 1 trials for hepatic metastases (HM) using intra-arterial (IA) administration (NCT01437007). Ongoing studies include hepatocellular carcinoma (HC) in phase 1/2, with active recruitment (NCT02191878), as well as completed phase 1/2 trials for neuroendocrine tumors (NET) and adrenocortical carcinoma (ACC) (NCT01262235). Another example is Atu027, developed by Silence Therapeutics GmbH, which has completed phase 1 testing for advanced solid cancer (ASC) using intravenous (IV) administration (NCT00938574). Similarly, ALN-TTR02, produced by Alnylam Pharmaceuticals for transthyretin amyloidosis (TTR-A), has completed phase 2 studies with IV administration (NCT01617967).

In addition, DOTAP-Chol-fus1 from the MD Anderson Cancer Center uses PLGA-based nanoparticles for lung cancer (LC) and has completed phase 1 testing with IV administration (NCT00059605). DCR-MYC, by Dicerna Pharmaceuticals, is being tested for solid tumors (ST), multiple myeloma (MM), and non–Hodgkin lymphoma (NHL). It is actively recruiting for both phase 1 (NCT02110563) and 1/2 (NCT02314052) studies, including hepatocellular carcinoma (HC) as one of the target conditions. PLGA-based nanoparticles are also used in siG12D LODER, a product of Silenseed, which is in phase 2 trials for pancreatic cancer (PC) via local administration (NCT01676259). Furthermore, Nitto Denko Corporation is conducting phase 1 studies for extensive hepatic fibrosis (EHF) with ND-L02-s0201 injection through IV administration (NCT02227459).

Continuing from the previous discussion, the successful performance of CALAA-01 in clinical trials has encouraged further investigation into nanoparticle-based gene delivery systems. Researchers are now examining different modifications and formulations in order to improve their efficiency and achieve more precise delivery to target sites. One important strategy is the optimization of nanoparticle composition to provide stronger stability and controlled release of therapeutic genes. In addition, researchers are attempting to refine targeting ligands so that they bind specifically to cancer cells, thus reducing off-target effects and improving the overall accuracy of treatment (Figure 6). Progress in nanotechnology has also supported the design of multifunctional nanoparticles that can transport several therapeutic agents at the same time. These complex systems are able to combine gene therapy with other treatment methods, including chemotherapy and immunotherapy, creating synergistic effects and improving therapeutic results. Such integration is highly valuable for addressing the complexity and heterogeneity of cancer, leading to more effective and individualized treatment options.

Figure 6

Figure 6. Proposed mechanism for CALAA-01. (a) Nanoparticles formed through the assembly of a linear polymer containing cyclodextrin (CDP), an adamantane-PEG conjugate (AD-PEG), a targeting element (transferrin, Tf), and the therapeutic gene (siRNA). (b) Patients receive an infusion of these nanoparticles. (c) Nanoparticles circulate and infiltrate tumors. (d) Receptor-mediated endocytosis occurs. (e) Targeted nanoparticles interact with receptors on the cancer cell surface. Reprinted from Davis (2009) with permission from American Chemical Society.

In recent years, investigators have also studied how the unique physicochemical features of nanoparticles may help overcome barriers in gene delivery, including the challenge of crossing the blood–brain barrier. This development has opened new opportunities for the treatment of neurological diseases through the targeted transport of therapeutic genes into specific regions of the brain. The adaptability of nanoparticles also makes it possible to design systems according to the needs of different diseases; this supports more tailored and efficient forms of gene therapy. As this field continues to expand, ongoing studies are working to resolve critical issues such as long-term safety, scalability, and production on a large scale. Cooperative work between researchers, clinicians, and industry representatives will play an essential role in turning these advances into practical, safe, and widely available treatments. The transition from laboratory research to clinical application of nanoparticle-based gene delivery systems shows great promise for transforming gene therapy and influencing the treatment of many diseases, particularly cancer. Table 5 provides an overview of the progress of clinical developments concerning nanoparticles used for gene delivery.

Table 5

Table 5. Clinical development of nanoparticles for gene delivery.

7 Challenges and regulatory considerations

Nanomaterials in gene therapy present several challenges that must be carefully considered (Duan et al., 2021). A major difficulty is the possible toxicity of some nanomaterials (Dai et al., 2021). Because they possess unique physicochemical properties, nanoparticles may interact in unintended ways with biological systems, producing cytotoxic or immunogenic effects (Soroush et al., 2016). Understanding the toxicological profile of nanomaterials is therefore essential to guarantee the safety of gene therapy applications (QIAO et al., 2021). Another continuing challenge is the delivery efficiency of nanomaterials to the target cells (Chen et al., 2017). Achieving accurate delivery while reducing off-target effects is a complex process (Liang et al., 2023). The creation of efficient delivery systems that can pass biological barriers and reach the desired site of action remains a critical issue in nanomedicine (Sheikhzadeh et al., 2021). Moreover, the long-term stability of nanomaterials and their possible accumulation in tissues or organs require careful investigation (He and Zhao, 2020). This involves the study of biodistribution and clearance pathways to prevent adverse consequences linked with prolonged exposure (Ojha et al., 2023).

Regulatory aspects play a key role in ensuring the safe and ethical application of nanomaterials in gene therapy (Tay and Melosh, 2019). Authorities such as the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) provide frameworks to evaluate the safety and effectiveness of therapies based on nanomaterials (Ghormade et al., 2011). Before starting clinical trials, researchers must satisfy regulatory requirements and submit detailed information on physicochemical properties, toxicity data, and manufacturing protocols (Shen Y. et al., 2018). This process ensures that potential risks and benefits are fully examined before human studies begin (Li et al., 2017). Ethical issues are also central to the regulatory environment (Kang et al., 2017). Studies involving human participants must go through strict ethical reviews to protect the health and rights of subjects (Yang et al., 2023). This requires obtaining informed consent, maintaining privacy, and guaranteeing transparency in research activities (Khaliq et al., 2023). Post-market surveillance and continuous monitoring are further important components of regulation (Han et al., 2021). Ongoing assessments after approval allow the detection of new safety problems and enable agencies to respond to protect public health (Szczeszak et al., 2015).

Addressing off-target effects in nanomaterial-based gene therapy requires multiple strategies (Agapito-Tenfen, 2016). One important method is designing nanocarriers with better specificity (Vélez et al., 2017). Surface modifications of nanoparticles can be used to improve their binding to target cells and reduce non-specific interactions (Zia et al., 2021). Advanced imaging technologies are also necessary to monitor biodistribution and pharmacokinetics of nanomaterials in real time (Flores et al., 2012). These techniques allow the detection of off-target accumulation and enable adjustments to improve delivery accuracy (Kapoor et al., 2018). The addition of intelligent targeting systems, such as ligands or antibodies, can further improve selectivity for certain cell types (Roma-rodrigues et al., 2020). These molecules bind to receptors on target cells, supporting accurate delivery and lowering off-target risks (Kumar et al., 2021). Furthermore, controlling the size and shape of nanomaterials can influence circulation time and uptake by cells (Haque et al., 2021). Adjusting these factors improves nanocarrier performance and reduces unwanted interactions (Salman et al., 2022). Preclinical investigations, both in vitro and in vivo, are necessary to assess safety and efficacy (Liang et al., 2023). These studies allow researchers to recognize off-target effects at early stages (Yang et al., 2015).

Reducing immunogenic responses in nanomaterial-based gene therapy requires a deep understanding of how nanoparticles interact with the immune system (Wang et al., 2018). One possible approach is to design nanomaterials with immunomodulatory characteristics to lower inflammation and encourage immune tolerance (Shen et al., 2021). Modifying the surface of nanocarriers can also decrease immune recognition (Pillai, 2014). For instance, PEGylation, in which nanoparticles are coated with polyethylene glycol, lowers immunogenicity and increases circulation time (Yang et al., 2015). Choosing biocompatible materials such as lipids or biodegradable polymers is equally important (Shen et al., 2022). Such materials are tolerated by the immune system and can limit harmful reactions (Pezoa et al., 2022). Comprehensive immunotoxicity testing in preclinical studies is essential to detect risks and develop effective countermeasures (Feng et al., 2023). Such evaluations include the effects of nanomaterials on immune cells, cytokine release, and immune balance (Fiaz et al., 2021). Finally, applying immune evasion methods, such as camouflage or combining with immunosuppressive agents, can improve the stealth features of nanomaterials and decrease immune detection (Samarasinghe et al., 2012).

The ethical application of nanomaterials in gene therapy requires careful attention to several main principles (Tian et al., 2013). Informed consent is essential to ensure that participants in clinical trials have a full understanding of the intervention, including the possible risks and benefits (Doroftei et al., 2021). Clear and open communication is necessary to maintain trust and respect for the autonomy of participants (Lai et al., 2017). Another important ethical principle is the protection of privacy (Yuan et al., 2023). Researchers need to apply strong data security systems to protect sensitive information collected during gene therapy trials (Khan, 2019). The use of anonymized data and strict confidentiality rules helps respect the privacy rights of participants (Foley et al., 2022).

Equitable access to gene therapies is also a central ethical concern (Ferdous et al., 2022). It is important to ensure that the benefits of nanomaterial-based gene therapy are available to all populations without discrimination (Kavanagh and Green, 2022). This includes solving problems of cost, availability, and inclusion in clinical trials, which are necessary parts of an ethical framework (Edagwa et al., 2017). Researchers must also reflect on the wider social impacts of gene therapy (Zheng et al., 2021). Considering and reducing possible negative outcomes, such as genetic discrimination or greater social inequalities, is essential (Hur and Chung, 2021). Ethical frameworks should guide all decisions to reduce harm and support social justice (Yang TC. et al., 2020). Ongoing ethical oversight, from the early laboratory stage to monitoring after approval, is also necessary (Majidi et al., 2016). Ethical review boards and regulatory bodies have an important role in protecting ethical standards and ensuring the responsible use of nanomaterials in gene therapy (Wang et al., 2015).

It is also important to flexibly adapt regulatory systems to the rapid progress of nanomaterial-based therapies (Xu et al., 2022). One key method is creating interdisciplinary cooperation between regulatory bodies, scientists, industry partners, and ethicists (Pissuwan et al., 2011; Zhu et al., 2022; Shen L. et al., 2018). This cooperation allows knowledge sharing, real-time risk assessment, and a broader understanding of the field (Erel-Akbaba et al., 2021). The regular updating of regulatory guidelines is necessary for including new scientific evidence and technological progress (Lai et al., 2017). Monitoring scientific studies, joining international collaborations, and working with expert panels can inform regulators about the latest advances in nanomedicine and gene therapy (Lin et al., 2015). Special regulatory pathways for nanomaterial-based therapies can also make the approval process more efficient (Han et al., 2021). Recognizing the unique opportunities and risks of nanotechnology helps the design of specific regulations that encourage innovation while protecting public health (Casper et al., 2023). Furthermore, promoting openness and communication between regulators and industry is essential (Kausar, 2021). Open discussion supports information exchange, helps achieve compliance, and improves responses to new challenges in nanomaterial-based gene therapies (Corte et al., 2019).

Incorporating adaptive licensing strategies, including conditional approvals and post-marketing monitoring, can address the dynamic development of nanomedicine (Ferdous et al., 2022). These strategies create a structure for gathering real-world data, revising risk–benefit evaluations, and making decisions that reflect advancing scientific knowledge (Calos, 2017). International cooperation is necessary to build consistent regulations and guarantee the safe and ethical application of nanomaterial-based gene therapies (Haque et al., 2021). Regulatory authorities from different regions can collaborate to harmonize guidelines, exchange best practices, and jointly respond to the worldwide challenges of this new field (van de Wiel et al., 2017). An effective method is the formation of international working groups or task forces devoted to nanotechnology in gene therapy (Feng et al., 2018). These groups bring experts from various agencies to evaluate scientific progress, exchange information, and design shared frameworks for testing nanomaterial safety and effectiveness (Zych et al., 2018). Participation in global conferences and workshops on nanomedicine and gene therapy also ensures that regulators remain informed about scientific developments (Maity et al., 2023). Such platforms encourage cooperation, networking, and the creation of international partnerships that strengthen regulatory coordination (Agapito-Tenfen, 2016).

The use of shared terminology and definitions for nanomaterials in gene therapy is essential (Su et al., 2016). Harmonization improves clarity in communication between regulators, scientists, and industry representatives (Lamboro et al., 2021). Open data exchange and the sharing of research findings promote transparency and strengthen consistent regulatory decisions (Gogolev et al., 2021). Creating a shared database or repository for nanomaterial safety and effectiveness supports the development of international standards (Ansari et al., 2020). Close cooperation between industry and regulatory authorities is also necessary for smoother approval processes (Zhou et al., 2022). Establishing clear communication and constructive relations promotes the exchange of information, ensures compliance, and accelerates the approval of innovative therapies (Gao et al., 2017). Early and transparent communication with regulators is particularly important (Joun et al., 2022). Companies should actively seek advice and feedback during preclinical and clinical development (Rong et al., 2020). This process helps align study designs, outcomes, and expectations, thus reducing the risk of delays in approval (Ibrahim et al., 2022). Involvement in regulatory pilot projects on nanomaterials in gene therapy provides practical insights and helps refine specialized approaches (Turcheniuk and Mochalin, 2017a). Such initiatives allow regulators and industry to test and improve methods designed for nanotechnology (Foley et al., 2022).

Strong and transparent data generation is also vital (Ding et al., 2022). Industry actors must conduct broad preclinical and clinical research, presenting regulators with detailed evidence on safety, quality, and effectiveness (Ghormade et al., 2011). This practice builds trust in the review process and speeds approvals (Koupaei et al., 2019). A risk-based regulatory approach also ensures an efficient use of resources (Kozielski et al., 2014). By prioritizing actions according to the level of risk linked to different aspects of nanomaterial-based therapies, stakeholders can create a more focused regulatory plan (Morán et al., 2018). At the same time, the development of international ethical standards for nanomaterials in gene therapy requires cooperation from global stakeholders (Wang et al., 2017). Agreement on ethical principles should involve experts, regulators, ethicists, researchers, and industry (Liang et al., 2023). Organizations such as the World Health Organization (WHO) and UNESCO may lead these efforts by coordinating ethical guidelines (Liu et al., 2018). Expert panels and working groups can specifically address the ethical dimensions of nanomaterial-based gene therapies to ensure inclusive solutions (Ferdous et al., 2022).

Public dialogue and engagement remain critical for including diverse perspectives in ethical standards (Edagwa et al., 2017). Ethical questions in gene therapy go beyond scientific or regulatory concerns since they involve social values and cultural contexts (Calos, 2017). Public consultations increase transparency and ensure that guidelines reflect global perspectives (Haque et al., 2021). A continuous review system is needed to respond to scientific and societal change (Feng et al., 2018). Updating ethical standards regularly guarantees their relevance and suitability for new developments (Turcheniuk and Mochalin, 2017a). International collaboration with bioethics organizations can also strengthen expertise and guidance (Rong et al., 2020). Aligning ethical frameworks with existing agreements and conventions helps create a unified global approach to the responsible use of nanomaterials in gene therapy (Shen et al., 2021).

8 Outlook and future research directions

Recent developments in nanomaterials for gene therapy focus on using different nanoparticles, such as liposomes, polymeric nanoparticles, and inorganic nanoparticles, to deliver genetic material into target cells (Flores et al., 2012). These nanomaterials act as carriers for therapeutic genes, supporting targeted and regulated gene expression (Gogolev et al., 2021). Liposomes are widely used because of their biocompatibility and ability to enclose both hydrophobic and hydrophilic gene loads (Kavanagh and Green, 2022). Polymeric nanoparticles provide design flexibility and allow precise control over release patterns (Haque et al., 2021). Inorganic nanoparticles, including gold and silica, are studied for their special features, such as surface modification to improve targeting and regulate release (Demirer et al., 2021). The field of nanomaterials in gene therapy is advancing quickly due to progress in materials science and biotechnology (Kaur et al., 2023). Current studies aim to create new materials with higher biocompatibility, lower toxicity, and better stability (Padayachee and Singh, 2020). Smart materials, including stimulus-responsive polymers, enable the controlled release of genetic material in reaction to specific physiological signals, improving therapeutic outcomes (Maity et al., 2023). Multifunctional nanomaterials that combine diagnostic and therapeutic functions are also attracting attention, supporting more precise and personalized gene therapy (Gad et al., 2020). Future directions in this area include innovative delivery strategies, such as cell-specific targeting and tissue-specific accumulation (Su et al., 2016). Developing nanomaterials for oral gene delivery is another important focus; it aims to solve difficulties linked to systemic administration (Xie et al., 2021). Furthermore, combining advanced imaging technologies with nanomaterials may allow the real-time monitoring of gene delivery, giving deeper insights into treatment effectiveness and possible side effects (Gogolev et al., 2021). These new trends in nanomaterials strongly influence gene therapy by improving accuracy, efficiency, and safety (Ansari et al., 2020). Creating nanomaterials with better pharmacokinetics and lower immune response improves the overall success of therapy (Zhou et al., 2022). Designing nanocarriers with particular properties, such as surface charge and size, also affects their interactions with biological barriers, influencing outcomes (Salman et al., 2022). The expected impact of nanomaterials in gene therapy in the near future is significant (Živojević et al., 2021). Nanocarriers may solve existing barriers, such as cell uptake, stability, and immune reactions (Song et al., 2023). Personalized nanotherapeutics that fit individual genetic profiles may soon become possible, opening the way to more targeted and effective treatments (Ibrahim et al., 2022). As the field grows, applying artificial intelligence (AI) and machine learning to improve delivery systems is expected to increase precision and efficacy (Miller and Siegwart, 2018). The progress of nanomaterials in gene therapy shows great potential to transform the field (Lastochkina et al., 2022). Present trends highlight the diversity of nanomaterials, from liposomes to inorganic nanoparticles, used in the controlled delivery of therapeutic genes (Wu K. et al., 2022). The continuous progress of nanomaterials, with better compatibility, reduced toxicity, and creative design, demonstrates the dynamic nature of this research (Xi et al., 2022). Future studies will likely emphasize advanced strategies, including oral delivery and cell-specific targeting, to address current challenges (Shen et al., 2021). New directions not only improve efficiency but also support multifunctional nanomaterials with diagnostic features (Kozielski et al., 2014). Looking ahead, nanomaterials are expected to bring transformative changes to gene therapy, making personalized treatment and AI-driven optimization possible (Mohamad et al., 2023). It is clear that nanomaterials will play a central role in shaping the future of gene therapy, with effects reaching far beyond the present state of the art (Wang et al., 2017).

Author contributions

BW: Writing – original draft, Writing – review and editing. JL: Writing – review and editing, Writing – original draft, Methodology. XZ: Methodology, Investigation, Project administration, Writing – original draft. RH: Supervision, Investigation, Methodology, Validation, Writing – review and editing, Data curation, Software, Visualization, Formal Analysis, Conceptualization, Resources, Writing – original draft, Funding acquisition, Project administration. HM: Writing – original draft, Conceptualization, Software, Funding acquisition, Resources, Writing – review and editing, Visualization, Investigation, Project administration, Methodology, Validation, Supervision, Formal Analysis, Data curation.

Funding

The authors declare that no financial support was received for the research and/or publication of this article.

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Generative AI statement

The author(s) declare that Generative AI was used in the creation of this manuscript. Generative AI was used in the preparation of this manuscript. Specifically, ChatGPT (OpenAI) was employed solely for language editing purposes. Its use was limited to suggesting alternative phrasings and improving clarity in certain non-technical sentences of the Introduction and abstract. No generative AI systems were used for data generation, analysis, interpretation, figures, or citations. All scientific content was created and validated by the authors, who assume full responsibility for the accuracy and integrity of the work.

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