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5 基于esgRNA/Cas9-RNP的ZmWx基因编辑突变体检测 A: 32个ZmWx的基因编辑材料的esgRNA/Cas9-RNP酶切检测; B: Sanger测序峰图。WT/WT为野生型, -1 bp/-1 bp为缺失1 bp的纯合突变体, WT/-3 bp为一个等位基因缺失3 bp的杂合突变体; CK: 未加esgRNA和Cas蛋白的ZmWx野生型PCR产物。

Fig. 5 Genotyping genome-edited ZmWx mutant via esgRNA/Cas9-RNP A: Genotyping genome-edited ZmWx mutant via esgRNA/Cas9-RNP; B: Sanger sequencing peak graph of different genotypes. WT/WT is a wild type, -1 bp/-1 bp is a homozygous mutant with 1 bp deleted, and WT/-3 bp is a heterozygous mutant with 3 bp allele deleted; CK: ZmWx wild PCR product without esgRNA and Cas protein.