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3 GhU6-5P::AE-sgRNA靶向突变的检测 A: M: 2K plus II DNA marker; 12、11分别为对照载体基因组PCR扩增产物酶切前后结果; 3~12为转化GhU6-5P::AE-sgRNA-Cas9单株样品基因组PCR扩增产物酶切后结果。B: 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; 3~12为转化GhU6-5P::AE-sgRNA-1300单株样品基因组PCR扩增产物酶切后结果。PCR/RE表示PCR扩增产物酶切分析。

Fig. 3 GhU6-5P::AE-sgRNA targeted mutations A: M: 2K plus II DNA marker; 12, 11: PCR/RE assay of negative control genome before and after the enzyme digestion; 3-12: PCR/RE assay of GhU6-5P::AE-sgRNA-Cas9 of single plant sample after enzyme digestion. B: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion; 3-12: PCR/RE assay of GhU6-5P::AE-sgRNA-1300 of single plant sample genome after enzyme digestion. PCR/RE indicates PCR/restriction enzyme assays.