An inducible CRISPR activation tool for accelerating plant regeneration

作者：Cuimei Zhang, Yajun Tang, Shanjie Tang, Lei Chen, Tong Li, Haidi Yuan, Yujun Xu, Yangyan Zhou, Shuaibin Zhang, Jianli Wang, Hongyu Wen, Wenbo Jiang, Yongzhen Pang, Xian Deng, Xiaofeng Cao, Junhui Zhou, Xianwei Song, Qikun Liu · 发表于：Plant Communications · 年份：2024 · DOI：10.1016/j.xplc.2024.100823 · 被引用次数：35 · 研究领域：Plant tissue culture and regeneration、CRISPR and Genetic Engineering、Plant Virus Research Studies

The inducible CRISPR activation (CRISPR-a) system offers unparalleled precision and versatility for regulating endogenous genes, making it highly sought after in plant research. In this study, we developed a chemically inducible CRISPR-a tool for plants called ER-Tag by combining the LexA-VP16-ER inducible system with the SunTag CRISPR-a system. We systematically compared different induction strategies and achieved high efficiency in target gene activation. We demonstrated that guide RNAs can be multiplexed and pooled for large-scale screening of effective morphogenic genes and gene pairs involved in plant regeneration. Further experiments showed that induced activation of these morphogenic genes can accelerate regeneration and improve regeneration efficiency in both eudicot and monocot plants, including alfalfa, woodland strawberry, and sheepgrass. Our study expands the CRISPR toolset in plants and provides a powerful new strategy for studying gene function when constitutive expression is not feasible or ideal.