Technological breakthroughs in generating transgene-free and genetically stable CRISPR-edited plants

作     者：Yubing He Yunde Zhao Yubing He;Yunde Zhao

作者机构：National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research(Wuhan)Huazhong Agricultural UniversityWuhan 430070China Section of Cell and Developmental BiologyUniversity of California San DiegoLa JollaCA 92093-0116USA 

出 版 物：《aBIOTECH》 (生物技术通报（英文版）)

年 卷 期：2020年第1卷第1期

页      面：88-96页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学] 0836[工学-生物工程] 

基　　金：This work was supported by grants from the National Transgenic Science and Technology Program(2019ZX08010-003 2019ZX08010-001)to YH. 

主　　题：CRISPR Transgene-free Marker-assisted selection TKC Cas9 gene editing 

摘      要：CRISPR/Cas9 gene-editing technologies have been very effective in editing target genes in all major crop plants and offer unprecedented potentials in crop improvement.A major challenge in using CRISPR gene-editing technology for agricultural applications is that the target gene-edited crop plants need to be transgene free to maintain trait stability and to gain regulatory approval for commercial production.In this article,we present various strategies for generating transgene-free and target geneedited crop plants.The CRISPR transgenes can be removed by genetic segregation if the crop plants are reproduced sexually.Marker-assisted tracking and eliminating transgenes greatly decrease the time and labor needed for identifying the ideal transgene-free plants.Transgenes can be programed to undergo self-elimination when CRISPR genes and suicide genes are sequentially activated,greatly accelerating the isolation of transgene-free and target gene-edited plants.Transgene-free plants can also be generated using approaches that are considered non-transgenic such as ribonucleoprotein transfection,transient expression of transgenes without DNA integration,and nano-biotechnology.Here,we discuss the advantages and disadvantages of the various strategies in generating transgene-free plants and provide guidance for adopting the best strategies in editing a crop plant.