HOXD基因簇内一系列CTCF位点反转揭示绝缘子功能

何象龙, 李金环, 吴强

Combinatorial CRISPR inversions of CTCF sites in HOXD cluster reveal complex insulator function

He Xianglong, Li Jinhuan, Wu Qiang

表1 引物序列

Table 1 Primers used in this study

类型 引物名称 序列(5′→3′) sgRNA a-sgRNA1F accgCGAAGAGTGCGGGAGAACGG a-sgRNA1R aaacCCGTTCTCCCGCACTCTTCG b-sgRNA1F accgGGCGCATCAGGAATGTAAG b-sgRNA1R aaacCTTACATTCCTGATGCGCC b-sgRNA2F accgTTCCAGAGATTATGAGCCAG b-sgRNA2R aaacCTGGCTCATAATCTCTGGAA c-sgRNA2F accgAGTAAAACTGGGTGTTGCC c-sgRNA2R aaacGGCAACACCCAGTTTTACT e-sgRNA1F accgAACTGTGCTCAAACGCTCTC e-sgRNA1R aaacGAGAGCGTTTGAGCACAGTT e-sgRNA2F accgGAGGCGCAAACAGCTGTTGT e-sgRNA2R aaacACAACAGCTGTTTGCGCCTC PCR a-1F TTCCAGCACCTCGGCTTTGTC a-1R CCCACTTTCCACCTCTGTCCTG b-1F GTCCGCCCGTGAGCTTCTGAA b-1R GCGTTGGCAGGGCTGGACTC b-1F1 TTCCCTGACCCTCCAAGCAC b-1R1 CTCACAGCAGCCGAAACCG b-2F GACACCTTGGTTCAGGCTCG b-2R CAGAGGCTGCATCAAACCAC c-1F TGATGCAGCCTCTGTGACCG c-1R AGTTTTCCCGTGGCGTCTGA c-2F GGACAACCCCTCACCTTGAA c-2R TATGGTGCCCAAAGTCCCAC e-1F TTCCCTGTCCCAGCTTGATTTC e-1R TCAACAGTGAAGGGCGGTGC e-2F CAAGCCACTCTCCCGCCACTA e-2R TCGCTCTCGTCCTCTCTTGGG bio-primer upstream-a-bioprimer 5′biotin-GGACGAGGGACATAGAGAGTTC downstream-e-bioprimer 5′biotin- AGGAGCCCAGGCATAGAGAC CS38-bioprimer 5′biotin-CCGAATTAAATCCCCGTGAC Island5-bioprimer 5′biotin-AAACACAAATGCATCAACCTG P5 primer upstream-a-P5 AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTCTGTCTAGAGATTCAAGTATTTCCCA P5 primer downstream-e-P5 AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTACGCGATACTCAAGGGAATACAAGCC CS38-P5 AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTAGGTTACAGTGTCATTTTCCCTG Island5-P5 AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTATGCATCTCATGAAGCTGGCATCT P7 index P7-index-1 CAAGCAGAAGACGGCATACGAGATCGAGTAATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-2 CAAGCAGAAGACGGCATACGAGATTCTCCGGAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-3 CAAGCAGAAGACGGCATACGAGATAATGAGCGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-4 CAAGCAGAAGACGGCATACGAGATGGAATCTCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-5 CAAGCAGAAGACGGCATACGAGATTTCTGAATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-6 CAAGCAGAAGACGGCATACGAGATACGAATTCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-7 CAAGCAGAAGACGGCATACGAGATAGCTTCAGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-8 CAAGCAGAAGACGGCATACGAGATGCGCATTAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-9 CAAGCAGAAGACGGCATACGAGATCATAGCCGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-10 CAAGCAGAAGACGGCATACGAGATTTCGCGGAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-11 CAAGCAGAAGACGGCATACGAGATGCGCGAGAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-12 CAAGCAGAAGACGGCATACGAGATCTATCGCTGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-13 CAAGCAGAAGACGGCATACGAGATAGAGTACTGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-14 CAAGCAGAAGACGGCATACGAGATGCTCCGTAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-15 CAAGCAGAAGACGGCATACGAGATCATGAGAGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-16 CAAGCAGAAGACGGCATACGAGATTGAATCGCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-17 CAAGCAGAAGACGGCATACGAGATGTCTGACTGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-18 CAAGCAGAAGACGGCATACGAGATCTGAATGCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-19 CAAGCAGAAGACGGCATACGAGATCGCTTCTGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-20 CAAGCAGAAGACGGCATACGAGATTCGCATGAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-21 CAAGCAGAAGACGGCATACGAGATAATAGCAGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-22 CAAGCAGAAGACGGCATACGAGATGTCGCGTAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-23 CAAGCAGAAGACGGCATACGAGATACGCGATAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7 index P7-index-24 CAAGCAGAAGACGGCATACGAGATTGATCGATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-25 CAAGCAGAAGACGGCATACGAGATCCGCATGAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-26 CAAGCAGAAGACGGCATACGAGATCCACAATCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-27 CAAGCAGAAGACGGCATACGAGATGATGTTCGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-28 CAAGCAGAAGACGGCATACGAGATGAATACGTGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-29 CAAGCAGAAGACGGCATACGAGATTAGATACCGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT P7-index-30 CAAGCAGAAGACGGCATACGAGATTATGGTCGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT adapter adapter-upper GACGTGTGCTCTTCCGATCTGNNNNNN-3`NH2 C6 adapter-lower 5′P-CAGATCGGAAGAGCACACGTC-3`NH2 C6