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1 基因编辑载体的酶切鉴定 A: 1、2分别为GhU6-5P::MAPKKK2-sgRNA-1300和GhU6-5P::MAPKKK2-sgRNA-Cas9. B: 1、2分别为GhU6-5P::AE-sgRNA-1300和GhU6-5P::AE-sgRNA-Cas9. C: 1~4分别为GhU6-5P-2::PDS-sgRNA-ClCrVA、GhU6-5P-2::CLA1-sgRNA-ClCrVA GhU6-5P-2::MAPKKK2- sgRNA-ClCrVA和GhU6-5P-2::AE-sgRNA-ClCrVA. M: 2K plus II DNA marker。

Fig. 1 Identification of gene editing vector by restriction enzyme digestion A: 1, 2 are GhU6-5P::MAPKKK2-sgRNA-1300 and GhU6-5P::MAPKKK2-sgRNA-Cas9. B: 1, 2 are GhU6-5P::AE-sgRNA-1300 and GhU6-5P:: AE-sgRNA-Cas9. C: 1-4 are GhU6-5P-2::PDS-sgRNA-ClCrVA , GhU6-5P-2::CLA1-sgRNA-ClCrVA, GhU6-5P-2::MAPKKK2-sgRNA- ClCrVA and GhU6-5P-2::AE-sgRNA-ClCrVA. M: 2K plus II DNA marker.