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10 海岛棉脱靶目标序列突变检测 A: OTtest-SG1-2引物检测; 1为转化Shift GbU6-5P-GGB- sgRNA1-Cas9I编辑载体, 2为Shift GbU6-5P-GGB-GbU6-4P- sgRNA1-ERA1-sgRNA2-Cas9I编辑载体, 3为转化Shift GbU6- 5P::GGB-sgRNA1-GbU6-4P::GGB-sgRNA2-Cas9I编辑载体。 B: OTtest-G1-2引物检测; 1为转化GbU6-5P-GGB-sgRNA1-Cas9I编辑载体, 2为GbU6-5P-GGB-GbU6-4P-sgRNA1-ERA1-sgRNA2- Cas9I编辑载体, 3为转化 GbU6-5P::GGB-sgRNA1-GbU6-4P:: GGB-sgRNA2-Cas9I编辑载体。C: OTtest-SG1-1引物检测; 1为转化Shift GbU6-5P-GGB-sgRNA1-Cas9I编辑载体, 2为Shift GbU6-5P-GGB-GbU6-4P-sgRNA1-ERA1-sgRNA2-Cas9I编辑载体, 3为转化Shift GbU6-5P::GGB-sgRNA1-GbU6-4P::GGB-sgRNA2- Cas9I编辑载体。

Fig. 10 Off target sequence mutation detection of Gossypium barbadense L. A: OTtest-SG1-2 primer detection; 1 is transformed Shift GbU6- 5P-GGB-sgRNA1-Cas9I edit vector, 2 is transformed Shift GbU6- 5P-GGB-sgRNA1-GbU6-4P::ERA1-sgRNA1-Cas9I edit vector, 3 is transformed Shift GbU6-5P-GGB-sgRNA1-GbU6-4P::GGB-sgRNA2- Cas9I edit vector. B:OTtest-G1-2 primer detection; 1 is transformed GbU6-5P-GGB-sgRNA1-Cas9I edit vector, 2 is transformed GbU6- 5P-GGB-sgRNA1-GbU6-4P::ERA1-sgRNA1-Cas9I edit vector, 3 is transformed GbU6-5P-GGB-sgRNA1-GbU6-4P::GGB-sgRNA2-Cas9I edit vector. C: OTtest-SG1-2 primer detection; 1 is transformed Shift GbU6-5P-GGB-sgRNA1-Cas9I edit vector, 2 is transformed Shift GbU6-5P-GGB-sgRNA1-GbU6-4P::ERA1-sgRNA1-Cas9I edit vector, 3 is transformed Shift GbU6-5P-GGB-sgRNA1-GbU6-4P::GGB-sgRNA2- Cas9I ditvector.