基于CRISPR/Cas9核糖核蛋白体DNA定点内切酶体外活性建立高效基因型分析技术

王南,祁显涛,刘昌林,谢传晓,朱金洁

Establishment of an efficient genotyping technique based on targeted DNA endonuclease in vitro activity of CRISPR/Cas9 ribonucleoprotein

Nan WANG,Xian-Tao QI,Chang-Lin LIU,Chuan-Xiao XIE,Jin-Jie ZHU

图4 基于esgRNA/Cas9NG-RNP检测体系对不同ZmWx位点的检测 A: Cas9-NG的6个新的靶向位点选择; B: 6个靶点对应的esgRNA 10% Urea-PAGE鉴定; C: PCR/RNP酶切分析; CK: 未加esgRNA和Cas蛋白的ZmWx野生型PCR产物。

Fig. 4 Cleavage assay of different targets for ZmWx locus via esgRNA/Cas9NG-RNP A: Targets selection of Cas9NG; B: 10% Urea-PAGE of 6 target-related esgRNA; C: PCR/RNP cleavage assay; CK: ZmWx wild PCR product without esgRNA and Cas protein.