摘要
靶向基因编辑系统是一种研究植物基因功能的工具。其中,CRISPR/Cas基因编辑系统的克隆策略相对简单,成本低廉,因而在生物学界获得了广泛的关注。为了更好地对其在植物中进行研究,笔者介绍了该系统的基本结构特征与作用机理,归纳了在拟南芥、烟草、水稻等植物中对该系统进行的报道,分析了该系统尚需改进的问题如脱靶效应,探讨了多种解决方法,如优化sg RNA序列长度、利用双Cas9切口酶、合成f Cas9复合体以及应用RFNs二聚体等,以降低脱靶效应。
Targeted gene editing system is a kind of tool to study the functions of plant genes. In this system,the cloning strategy of CRISPR/Cas gene editing system is relatively simple and the cost is low, thus it hasdrawn widespread attention in biology. In order to study the system in plant better, the author introduced thebasic structure and the mechanism of the system, summarized research reports on the system in plants likeArabidopsis, tobacco, rice and so on, analyzed problems of the system that needs to be improved, such as off-target effects, and discussed several solutions, including optimizing sg RNA sequence length, using doubleCas9 nickases, synthesizing f Cas9 complex and applying RFNs dimers, to reduce off-target effects.
出处
《中国农学通报》
2015年第12期187-192,共6页
Chinese Agricultural Science Bulletin
