棉花CRISPR/Cas9基因编辑有效sgRNA高效筛选体系的研究

周冠彤, 雷建峰, 代培红, 刘超, 李月, 刘晓东

Efficient screening system of effective sgRNA for cotton CRISPR/Cas9 gene editing

ZHOU Guan-Tong, LEI Jian-Feng, DAI Pei-Hong, LIU Chao, LI Yue, LIU Xiao-Dong

图4 GhU6-5P-2::MAPKKK2-sgRNA-CLCrVA部分编辑效应测序检测结果 A: M: 2K plus II DNA marker; 1、2分别为对照载体基因组PCR扩增产物酶切前后结果。B: M: 2K plus II DNA marker; 3~6为转化GhU6-5P-2::MAPKKK2-sgRNA-CLCrVA单株样品基因组PCR扩增产物酶切后结果。C: M: 2K plus II DNA marker; 1~10为单克隆菌液PCR产物酶切后结果。PCR/RE表示PCR扩增产物酶切分析。

Fig. 4 Results of GhU6-5P-2::MAPKKK2-sgRNA-CLCrVA partial editing effect sequencing A: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion. B: M: 2K plus II DNA marker; 3-6: PCR/RE assay of GhU6-5P::MAPKKK2-sgRNA-CLCrVA genome of single plant sample after enzyme digestion. C: M: 2K plus II DNA marker, 1-10: clony PCR product after enzyme digestion. PCR/RE indicates PCR/restriction enzyme assays.