锌指核酸酶技术在植物基因工程中的应用

锌指核酸酶(zinc finger nucleases,ZFNs)由3到4个锌指结构(zinc fingers,ZFs)和FokⅠ核酸内切酶的剪切结构域组成。锌指核酸酶(ZFNs)通过锌指结构(ZFs)与特异核酸位点结合,再利用FokⅠ的酶切作用切割DNA,引起特异位点DNA双链断裂(double strand break,DSB)。DNA双链断裂可以通过非同源末端连接(non-homologous end joining,NHEJ) 或同源重组(homologous recombination,HR)来修复。在修复过程中实现对基因组DNA的靶向修饰。介绍了锌指核酸酶结构、人工构建途径,作用机理和试验步骤,重点综述了锌指核酸酶技术在植物基因工程的应用。

Zinc Finger Nucleases and Targeted Genome Engineering in Plants

Zinc finger nucleases (ZFNs) typically consist of three to four zinc fingers (ZFs) and the non-specific DNA-cleavage domain of the restriction endonuclease FokⅠ. In this configuration, the ZFs constitute the binding module and the FokⅠ domain the cleavage module. ZFNs offer a general way to deliver a targeted site-specific double strand break (DSB) to the genome. DSBs are mainly repaired by two pathways, non-homologous end joining (NHEJ) and homologous recombination (HR), which targeted mutagenesis will be generated. In this review, we are first describe the structure of ZFNs, and ZFN-mediated targeted genome engineering via HR and NHEJ,and the methods of construct novel ZFNs and the steps of ZFN-mediated gene targeting. Then we mainly explore that ZFNs were used in plants.