BRIEF RESEARCH REPORT article
Front. Plant Sci.
Sec. Plant Biotechnology
Volume 16 - 2025 | doi: 10.3389/fpls.2025.1620874
This article is part of the Research TopicEngineering Future Crops Through Genome EditingView all 16 articles
A Simple and Efficient System for Evaluating Plant Genome Editing Efficiency and Its Application in Optimizing the ISAam1 TnpB Nuclease
Provisionally accepted
- 1Key Laboratory of Herbage and Endemic Crop Biology, Ministry of Education, Inner Mongolia University, Hohhot, China
- 2Inner Mongolia University, Hohhot, China
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Genome editing technology has revolutionized plant genetic breeding. However, Significant variability in editing activity has been observed across different genome editing systems and target sites, highlighting the importance of developing efficient evaluation systems for assessing genome editing efficiency in plants. In this study, we developed a simple, rapid, and efficient system based on hairy root transformation to evaluate somatic genome editing efficiency in plants. This system is easy to implement, does not require sterile conditions, and enables visual identification of transgenic hairy roots within two weeks. We first validated the system using the CRISPR/Cas9 genome editing platform, confirming its effectiveness. Subsequently, we applied this system to assess the somatic editing activity of the recently identified ISAam1 TnpB nucleases, which show considerable promise for plant genome editing applications. Furthermore, through protein engineering, we identified two variants, ISAam1(N3Y) and ISAam1(T296R), which exhibited a 5.1-fold and 4.4-fold enhancement in somatic editing efficiency, respectively. These findings demonstrate that the developed method provides an effective tool for optimizing genome editing system and screening potential target sites in plant genomes.
Keywords: Genome editing, Agrobacterium rhizogenes, Plants, ISAam1, TnpB;
Received: 30 Apr 2025; Accepted: 11 Aug 2025.
Copyright: © 2025 Cao, Bai, Li and Sun. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Yongwei Sun, Inner Mongolia University, Hohhot, China
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