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RNA interference (RNAi) created a global excitement around the turn of the century and small interfering RNA (siRNA), small hairpin RNA (shRNA), and micro RNAs (miRNA) were the center of attention for two decades to silence gene expression. Recent analysis shows a decline in the number of published studies ...

RNA interference (RNAi) created a global excitement around the turn of the century and small interfering RNA (siRNA), small hairpin RNA (shRNA), and micro RNAs (miRNA) were the center of attention for two decades to silence gene expression. Recent analysis shows a decline in the number of published studies focusing on these RNAi approaches since 2015. This could be partially explained by the difficulties with in vivo implementation of RNAi approach, which is mostly related to the challenges with effective delivery of functional nucleic acids. New developments with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 has created a similar excitement due to its potential to edit genes permanently, which is a distinct advantage over the temporary protein silencing achieved with RNAi. For the success of this approach, CRISPR RNA/trans-activating CRISPR RNA (tracrRNA) and Cas9 are required in the cell at the same time, which has brought on different delivery approaches.

Effective implementation of CRISPR has been pursued by delivery of a variety of combinations to target cells, including the complex of Cas9 protein and double-stranded crRNA/tracrRNA (known as ribonucleoproteins, or RNP), mRNA that expresses Cas9 in the target cell and double-stranded crRNA/tracrRNA, and a plasmid containing all the required nucleic acids together (including Cas9 nuclease expression cassette and a guide RNA cloning cassette). However, regardless of the specific approach, delivering the required CRISPR components into target cells faces many challenges that are shared with previously studied RNAi approach to protein silencing. An added complication is the delivery of multiple components with different physicochemical features. Recent promising results with CRISPR/Cas9 approach in a small population of patients with hereditary ATTR can open the doors to more effective clinical applications. A better understanding of different CRISPR/Cas9 approaches and challenges of effective delivery of CRISPR/Cas9 tools will enhance our chances of taking advantage of this powerful tool in translational and clinical research.


This collection aims to attract submissions focused (but not limited on) the following topics:

Different approaches to delivering CRISPR/Cas9 components

Recent accomplishments in CRISPR/Cas9 technology in animal models

Challenges for in vivo delivery of CRISPR/Cas9

CRISPR/Cas9 gene silencing vs. RNA interference

Synthetic vectors for delivery of CRISPR components

Viral vectors for delivery of CRISPR components


Note: Dr. Giedrius Gasiunas is an employee and co-founder of CasZyme.

Keywords: CRISPR/Cas9, Delivery, Vectors, Cancer, in vitro, Animal studies


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