CRISPR/Cas9介导的绵羊示踪脐带间充质干细胞系的建立

李松美,仇雨歌,陈胜男,王晓萌,王春生

CRISPR/Cas9 Mediated Exogenous Gene Knock-in at ROSA26 Locus in Sheep Umbilical Cord Mesenchymal Stem Cells

LI SongMei,QIU YuGe,CHEN ShengNan,WANG XiaoMeng,WANG ChunSheng

表1 sgRNA和引物及其目的片段大小

Table 1 Sequences and product length of sgRNA and primers

sgRNA/引物 sgRNA/Primers 序列 Oligonucleotide sequences 片段 Length (bp) KOD-R 5′-GGTGTTTCGTCCTTTCCAC 19 px330-sgRNA-1 5′-CCAGCAGGTATAAGATTTAGGTTTTAGAGCTAGAATAGCAGGT 43 px330-sgRNA-2 5′-CCTCTAAATCTTATACCTGCGTTTTAGAGCTAGAATAGCAGGT 43 px330-sgRNA-3 5′-TGTCCTGCAGTGGATCCAGCGTTTTAGAGCTAGAATAGCAGGT 43 px330-TP-F 5′-GCTGCCTGAAGGACAAGACTA 21 px330-TP-R 5′-GGCAACACCTGGGACTGATTT 21 sROSA26-LHA-F 5′-ACTAGTTACGCTGAAAGGGAAAGAGG 26 sROSA26-LHA-R 5′-ACGCGTAGGACAACGCCCAGGATT 24 sROSA26-RHA-F 5′-TGTACACCCAATTTCTTTATCTTCCC 26 sROSA26-RHA-R 5′-ACATGTTCCTGTCAGTAGTTACCACCC 27 SRL-F 5′-AAGAGGCTGTGCTCTGGG 18 SRL-R 5′-CGTGAGTCAAACCGCTATCC 20 SRR-F 5′-TCCCTAAAGAAACAGTGGC 19 SRR-R 5′-CACGTTTGTGATGATGGAAT 20