CRISPR-Cas9植物基因编辑系统敲除拟南芥AG基因表达载体的构建

The Construction of Expression Vector of Knocking Out AG Gene in Arabidopsis thaliana by CRISPR-Cas9

CRISPR-Cas系统是细菌和古细菌在进化过程中逐渐形成的一种适应性免疫系统,通过sg RNA介导对靶位点进行定位并利用Cas酶对核酸实现双链断裂(double-strand break,DSB)。CRISPR-Cas系统作为一种新兴的基因定点编辑技术逐渐成熟并在多个植物中成功得到应用,以此技术对目的基因进行靶向的敲除或敲入。本实验以拟南芥AG基因为基础,构建CRISPR-Cas9植物基因编辑系统敲除拟南芥AG基因的表达载体,并利用农杆菌介导CRISPR-Cas9植物基因编辑系统敲除拟南芥AG基因,以获得高效的敲除拟南芥基因的遗传转化体系。

The CRISPR-Cas system is a kind of adaptive immune system,which was gradually formed in the evolutionary process of bacteria and archaea.The CRISPR-Cas system positions to the targeted point with sg RNA,and the nucleic acid is double-strand brokenby Cas enzyme.CRISPR-Cas system as a new gene fixed-point editing techniques has been successful applied in many plants.Targeted genes can be targeting knock-out and knock-in by the technology of CRISPRs.The experiment is based on the AG gene of Arabidopsis thaliana,and construced a vector which knockout AG gene by CRISPR-Cas9 system,and then the high efficient genetic transformation system is obtained which can knockout Arabidopsis thaliana genes by using the agrobacterium-mediated genetic transformation of CRISPR-Cas9 system.