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9 海岛棉脱靶目标序列扩增检测 转化No shift sgRNA类型靶序列: 1、2分别为转化GbU6-5P-GGB-sgRNA1-Cas9I和对照编辑载体, 3、4为GbU6-5P- GGB-GbU6-4P-sgRNA1- ERA1-sgRNA2-Cas9I和对照编辑载体, 5、6为转化GbU6-5P::GGB-sgRNA1-GbU6-4P::GGB-sgRNA2 -Cas9I和对照编辑载体。转化Shift sgRNA类型靶序列: 1、2分别为转化Shift GbU6-5P-GGB-sgRNA1-Cas9I和对照编辑载体, 3、4为Shift GbU6-5P-GGB-GbU6-4P-sgRNA1-ERA1-sgRNA2- Cas9I和对照编辑载体, 5、6为转化Shift GbU6-5P::GGB- sgRNA1-GbU6-4P::GGB-sgRNA2-Cas9I和对照编辑载体。

Fig. 9 Off target sequence amplification test of Gossypium barbadense L. Transform No shift sgRNA type target sequence: 1 and 2 is transformed GbU6-5P-GGB-sgRNA1-Cas9I edit vector and Control edit vector, 3 and 4 is transformed GbU6-5P-GGB-sgRNA1-GbU6- 4P::ERA1-sgRNA1-Cas9I edit vector and Control edit vector, 5 and 6 is transformed GbU6-5P-GGB-sgRNA1-GbU6-4P::GGB-sgRNA2- Cas9I edit vector and Control edit vector. Transform Shift sgRNA type target sequence: 1 and 2 is transformed Shift GbU6-5P- GGB-sgRNA1-Cas9I edit vector and Control edit vector, 3 and 4 is transformed Shift GbU6-5P-GGB-sgRNA1-GbU6-4P::ERA1-sgRNA1- Cas9I edit vector and Control edit vector, 5 and 6 is transformed Shift GbU6-5P-GGB-sgRNA1-GbU6-4P::GGB-sgRNA2-Cas9I edit vector and Control edit vector.