基于优化sgRNA系统提高海岛棉CRISPR/Cas9基因组编辑功效的研究

李继洋,胡燕,姚瑞,代培红,刘晓东

Enhancing CRISPR/Cas9 genomic editing efficiency based on optimization of sgRNA of Gossypium barbadense L.

Ji-Yang LI,Yan HU,Rui YAO,Pei-Hong DAI,Xiao-Dong LIU

图6 海岛棉转化不同类型靶序列碱基突变测序结果 A: 转化GbU6-5P-GGB-sgRNA1-Cas9I载体靶位点编辑效应测序检测结果; B: 转化GbU6-5P::GGB-sgRNA1-GbU6-4P::ERA1- sgRNA2-Cas9I载体GGB靶位点编辑效应测序检测结果; C: 转化GbU6-5P::GGB-sgRNA1-GbU6-4P::ERA1-sgRNA2-Cas9I载体ERA1靶位点编辑效应测序检测结果; D: 转化GbU6-5P::GGB-sgRNA1-GbU6-4P::GGB-sgRNA2-Cas9I载体GGB-sgRNA2靶位点编辑效应测序检测结果; E: 转化GbU6-5P::GGB-sgRNA1-GbU6-4P::GGB-sgRNA2-Cas9I载体GGB-sgRNA1靶位点编辑效应测序检测结果。红色框为PAM位点的位置。

Fig. 6 Sequencing results of base mutation of different types of target sequences of Gossypium barbadense L. A: Transformation of GbU6-5P-GGB-sgRNA 1-Cas9I vector target site editing effect sequencing detection results; B: Transformation GbU6- 5P::GGB-sgRNA1-GbU6-4P::ERA1-sgRNA2-Cas9I vector GGB target site editing effect sequencing detection results; C: GbU6-5P::GGB- sgRNA1-GbU6-4P::ERA1-sgRNA2-Cas9I vector ERA1 target site editing sequencing results; D: transformation GbU6-5P::GGB-sgRNA1-GbU6- 4P::GGB-sgRNA2-Cas9I vector GGB-sgRNA2 target site editing effect sequencing test results; E: transformation GbU6-5P::GGB-sgRNA1-GbU6- 4P::GGB-sgRNA2-Cas9I vector GGB-sgRNA1 target site editing effect sequencing test results. The red box is the location of the PAM site.