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3 基于esgRNA /Cas9-RNP及esgRNA/ Cas9NG-RNP检测体系的优化 A: esgRNA梯度;B: Cas9蛋白梯度; C: 酶切反应时间梯度; D: 不同基因型PCR产物酶切; E: esgRNA梯度; F: Cas9NG蛋白梯度; G: 酶切反应时间梯度; H: 不同基因型PCR产物酶切; CK: 未加esgRNA和Cas蛋白的ZmWx 野生型PCR产物。

Fig. 3 Optimization of cleavage assay via esgRNA /Cas9-RNP and esgRNA/ Cas9NG-RNP A: Effect of different gradients of esgRNA; B: Effect of different gradients of Cas9 protein; C: Effect of different gradients of digestion time; D: Digestion of PCR products of different genotypes; E: Effect of different gradients of esgRNA; F: Effect of different gradients of Cas9NG protein; G: Effect of different gradients of digestion time; H: Digestion of PCR products of different genotypes; CK: ZmWx wild PCR product without esgRNA and Cas protein.