Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

作     者：HE Yubing ZHU Min WANG Lihao WU Junhua WANG Qiaoyan WANG Rongchen ZHAO Yunde 

作者机构：National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan) Huazhong Agricultural University College of Life Science and Technology Huazhong Agricultural University College of Plant Science & Technology Huazhong Agricultural University Section of Cell and Developmental Biology University of California San Diego 

出 版 物：《Rice science》 (水稻科学（英文版）)

年 卷 期：2019年第26卷第2期

页      面：109-117,I0016,I0017页

核心收录：

学科分类：09[农学] 

基　　金：supported by Chinese Ministry of Agriculture and Rural Affairs (Grant No. 2018ZX0801003B) the National Transgenic Science and Technology Program (Grant No. 2016ZX08010002) 

主　　题：genome editing suicide gene transgene killer CRISPR Cas9 transgene-free 

摘      要：Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR(TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35 S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.