Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy

Cheng, Yiqiu1,2; Zhang, Lei1,2; Li, Jing1,2; Dang, Xiaofei1; Zhu, Jian-Kang3,4,5; Shimada, Hiroaki6; Miki, Daisuke1

2024-03-13

10.3389/fpls.2024.1360925

FRONTIERS IN PLANT SCIENCE   影响因子和分区

1664-462X

Gene targeting (GT) is a promising tool for precise manipulation of genome sequences, however, GT in seed plants remains a challenging task. The simple and direct way to improve the efficiency of GT via homology-directed repair (HDR) is to increase the frequency of double-strand breaks (DSBs) at target sites in plants. Here we report an all-in-one approach of GT in Arabidopsis by combining a transcriptional and a translational enhancer for the Cas expression. We find that facilitating the expression of Cas9 and Cas12a variant by using enhancers can improve DSB and subsequent knock-in efficiency in the Arabidopsis genome. These results indicate that simply increasing Cas protein expression at specific timings - egg cells and early embryos - can improve the establishment of heritable GTs. This simple approach allows for routine genome engineering in plants.

genome engineering CRISPR/Cas9 CRISPR/ttCas12a RPS5a all-in-one gene targeting (GT) transcriptional and translational enhancers

SCI

英语

其他

Shanghai Science and Technology Innovation Plan["20ZR1467000","23WZ2500800"] ; Foreign Expert Project[G202201355L] ; Ministry of Education, Culture, Sports, Science and Technology (MEXT)[21570050] ; National Key R&D Program of China[2021YFA1300404] ; National Natural Science Foundation of China[32188102]

Plant Sciences

Plant Sciences

WOS:001190876100001

FRONTIERS MEDIA SA

Web of Science

1.Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, CAS Ctr Excellence Mol Plant Sci, Shanghai, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
3.Southern Univ Sci & Technol, Inst Adv Biotechnol, Shenzhen, Peoples R China
4.Southern Univ Sci & Technol, Sch Life Sci, Shenzhen, Peoples R China
5.Chinese Acad Agr Sci, Ctr Adv Bioind Technol, Beijing, Peoples R China
6.Tokyo Univ Sci, Dept Biol Sci & Technol, Tokyo, Japan

Cheng, Yiqiu,Zhang, Lei,Li, Jing,et al. Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy[J]. FRONTIERS IN PLANT SCIENCE,2024,15.

Cheng, Yiqiu.,Zhang, Lei.,Li, Jing.,Dang, Xiaofei.,Zhu, Jian-Kang.,...&Miki, Daisuke.(2024).Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy.FRONTIERS IN PLANT SCIENCE,15.

Cheng, Yiqiu,et al."Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy".FRONTIERS IN PLANT SCIENCE 15(2024).