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8 转化海岛棉不同类型靶序列的原生质体Cas9及对应sgRNA半定量检测结果 1、2分别为转化GbU6-5P-GGB-sgRNA1-Cas9I和对照编辑载体检测GGB-sgRNA1, 3、4为GbU6-5P-GGB-sgRNA1-GbU6-4P- ERA1-sgRNA2-Cas9I编辑载体分别检测GGB-sgRNA1、ERA1- sgRNA2, 5、6为转化GbU6-5P::GGB-sgRNA1-GbU6-4P::GGB- sgRNA2-Cas9I分别检测GGB-sgRNA1、GGB-sgRNA2, 7为转化与GbU6-5P-sgRNA1-4P-sgRNA1Cas9I对照编辑载体检测GGB- sgRNA1。

Fig. 8 Semi-quantitative detection of protoplast Cas9 and corresponding sgRNA in different types of target sequences transformed in Gossypium barbadense L. 1 and 2 is transformed GbU6-5P-GGB-sgRNA1-Cas9I edit vector and Control edit vector were detected GGB-sgRNA1, 3 and 4 is transformed GbU6-5P-GGB-sgRNA1-GbU6-4P::ERA1-sgRNA1- Cas9I edit vector were detected GGB-sgRNA1, ERA1-sgRNA2, 5 and 6 is transformed GbU6-5P-GGB-sgRNA1-GbU6-4P::GGB- sgRNA2-Cas9I edit vector were detected GGB-sgRNA1, GGB- sgRNA2, 7 is transformed GbU6-5P-sgRNA1-4P-sgRNA1-Cas9I control edit vector were detected GGB-sgRNA1.