通过农杆菌浸润法利用双生病毒将CRISPR/Cas9组件导入烟草
Use of Geminivirus for Delivery of CRISPR/Cas9 Components to Tobacco by Agro-infiltration.
作者信息
Yin Kangquan, Han Ting, Liu Yule
机构信息
Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China.
State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
出版信息
Bio Protoc. 2017 Apr 5;7(7):e2209. doi: 10.21769/BioProtoc.2209.
CRISPR/Cas9 system is a recently developed genome editing tool, and its power has been demonstrated in many organisms, including some plant species ( Wang , 2016 ). In eukaryotes, the Cas9/gRNA complexes target genome sites specifically and cleave them to produce double-strand breaks (DSBs), which can be repaired by non-homologous end joining (NHEJ) pathway ( Wang , 2016 ). Since NHEJ is error prone, mutations are thus generated. In plants, delivery of genome editing reagents is still challenging. In this protocol, we detail the procedure of a virus-based gRNA delivery system for CRISPR/Cas9 mediated plant genome editing (VIGE). This method offers a rapid and efficient way to deliver gRNA into plant cells, especially for those that are recalcitrant to transformation with .
CRISPR/Cas9系统是一种最近开发的基因组编辑工具，其功能已在许多生物体中得到证明，包括一些植物物种（Wang，2016）。在真核生物中，Cas9/gRNA复合物特异性靶向基因组位点并切割它们以产生双链断裂（DSB），这些双链断裂可通过非同源末端连接（NHEJ）途径修复（Wang，2016）。由于NHEJ容易出错，因此会产生突变。在植物中，基因组编辑试剂的递送仍然具有挑战性。在本方案中，我们详细介绍了一种基于病毒的gRNA递送系统用于CRISPR/Cas9介导的植物基因组编辑（VIGE）的程序。这种方法提供了一种将gRNA快速有效地递送到植物细胞中的方法，特别是对于那些难以通过转化的细胞。
Zotero 插件