Genome editing in plants using the TnpB transposase system

作者：Qi Li, Yongqiang Wang, Zhuoting Hou, Hang Zong, Xuping Wang, Yong E. Zhang, Haoyi Wang, Haitao Chen, Wen Wang, Kang Duan · 发表于：aBIOTECH · 年份：2024 · DOI：10.1007/s42994-024-00172-6 · 被引用次数：26 · 研究领域：CRISPR and Genetic Engineering、Chromosomal and Genetic Variations、RNA regulation and disease

The widely used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease (Cas) system is thought to have evolved from IS200/IS605 transposons. TnpB proteins, encoded by one type of IS200/IS605 transposon, are considered to be the evolutionary ancestors of Cas12 nucleases, which have been engineered to function as RNA-guided DNA endonucleases for genome editing in bacteria and human cells. TnpB nucleases, which are smaller than Cas nucleases, have been engineered for use in genome editing in animal systems, but the feasibility of this approach in plants remained unknown. Here, we obtained stably transformed genome-edited mutants in rice ( Oryza sativa ) by adapting three recently identified TnpB genome editing vectors, encoding distinct TnpB nucleases (ISAam1, ISDra2, and ISYmu1), for use in plants, demonstrating that the hypercompact TnpB proteins can effectively edit plant genomes. ISDra2 and ISYmu1 precisely edited their target sequences, with no off-target mutations detected, showing that TnpB transposon nucleases are suitable for development into a new genome editing tool for plants. Future modifications improving the genome-editing efficiency of the TnpB system will facilitate plant functional studies and breeding programs.