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

表1 本研究使用的引物信息

Table 1 Primers used in this study

序列名称 Sequence name 序列 Sequence (5'-3') PAM位点序列 PAM site sequence Shift GbU6-4PERA1-sg2F GATTGTCTTTCGCAGAATGCATGA CGG Shift GbU6-4PERA1-sg2R AAACTCATGCATTCTGCGAAAGAC GbU6-4PERA1-sg2F GATTGTTCGCAGAATGCATGACGG TGG GbU6-4PERA1-sg2R AAACCCGTCATGCATTCTGCGAAC GbU6-5PGGB-sg1F AAGTGTGTCGAAGTACTGAAGCGG CGG GbU6-5PGGB-sg1R AAACCCGCTTCAGTACTTCGACAC GbU6-4PGGB-sg2F GATTGTCTTTCAGTCTTATGATGG TGG GbU6-4PGGB-sg2R AAACCCATCATAAGACTGAAAGAC Shift GbU6-5PGGB-sg1F AAGTGCTCTGTCGAAGTACTGAAG CGG Shift GbU6-5PGGB-sg1R AAACCTTCAGTACTTCGACAGAGC Shift GbU6-4PGGB-sg2F GATTGTGTTCTTTCAGTCTTATGA TGG Shift GbU6-4PGGB-sg2R AAACTCATAAGACTGAAAGAACAC Test GGB-sg1F AAGTGGAAAGAGAATGGCGAC Test GGB-sg1R AGCTAATTCGCTATTGCAATCAATC Test GGB-sgR ACCATGTGATTCTAAACCAGG Test ERA1-sg1F TAATAGGTGGTCAGGTTATGC Test ERA1-sg1R AGGAGACTTGCAACCTGATC FJ-1F GTAAAACGACGGCCAG FJ-1R CAGGAAACAGCTATGAC FJ-2F TAAACTGAAGGCGGGAAACG FJ-2R CGGTTCTGTCAGTTCCAAACG