[发明专利]一类调控CCR5和CXCR4基因的融合蛋白及方法

权利要求书

1.一种类转录激活因子(Transcription Activator Like Effectors，TALE)，其特征在于所述类转录激活因子(TALE)能够与人类CCR5基因活性区域或者CXCR4基因活性区域中的DNA片段识别并结合，所述人类CCR5基因活性区域一共有3个，其核苷酸序列分别为：活性区域1)：tgcacagggtggaacaagatggattatcaagtgtcaagtccaatctatgacatcaattattatacatcggagccctgccaaaaaatcaatgtgaagcaaatcgcag，活性区域2)：caacatgctggtcatcctcatcctgataaactgcaaaaggctgaagagcatgactgacatctacctg，活性区域3)：tcttcattacacctgcagctctcattttccatacagtcagtatcaattctggaagaatttccagacattaaagatag；所述CXCR4基因活性区域一共有2个，其核苷酸序列分别为：活性区域1’)：TAACTACACCGAGGAAATGGGCTCAGGGGACTATGACTCCATGAAGGAACCCTGTTTCCGTGAAGAAAATGCTAATTTCAATAAAA，活性区域2’)：TAAAATCTTCCTGCCCACCATCTACTCCATCATCTTCTTAACTGGCATTGTGGGCAATGGATTGGTCATCCTGGTCATGGGTTACCAGAAGAAACTGAGAAGCATGACGGACAAGTACAGGCTGCA，若对CCR5或者CXCR4上述活性区域中的DNA片段进行切除，则待基因修复之后，HIV病毒将不能再攻击这些基因修复之后的细胞；

所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1)，其核苷酸序列为5’-TATGACATCAATTATTAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI NG NN NI HD NI NG HD NI NI NG NG NI NG NG NI NG-C端，上述类转录激活因子(TALE)记为“TALE-1-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1)的反义链，其核苷酸序列为5’-TGCTTCACATTGATTTTTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN HD NG NG HD NI HD NI NG NG NN NI NG NG NG NG NG NG-C端，上述类转录激活因子(TALE)记为“TALE-1-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1)，其核苷酸序列为5’-TGGAACAAGATGGATTAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN NN NI NI HD NI NI NN NI NG NN NN NI NG NG NI NG-C端，上述类转录激活因子(TALE)记为“TALE-2-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1)的反义链，其核苷酸序列为5’-TAATTGATGTCATAGAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI NI NG NG NN NI NG NN NG HD NI NG NI NN NI NG-C端，上述类转录激活因子(TALE)记为“TALE-2-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1)，其核苷酸序列为5’-TATCAAGTGTCAAGTCCAAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI NG HD NI NI NN NG NN NG HD NI NI NN NG HD HD NI NI NG-C端，上述类转录激活因子(TALE)记为“TALE-3-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1)的反义链，其核苷酸序列为5’-TGGCAGGGCTCCGATGTATAAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN NN HD NI NN NNNN HD NG HD HD NN NI NG NN NG NI NG NI NI NG-C端，上述类转录激活因子(TALE)记为“TALE-3-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2)，其核苷酸序列为5’-TGCTGGTCATCCTCAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN HD NG NN NN NG HD NI NG HD HD NG HD NI NG-C端，上述类转录激活因子(TALE)记为“TALE-4-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2)的反义链，其核苷酸序列为5’-TCATGCTCTTCAGCCTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-HD NI NG NN HD NG HD NG NG HD NI NN HD HD NG NG-C端，上述类转录激活因子(TALE)记为“TALE-4-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2)，其核苷酸序列为5’-TGGTCATCCTCATCCT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN NN NG HD NI NG HD HD NG HD NI NG HD HD NG-C端，上述类转录激活因子(TALE)记为“TALE-5-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2)的反义链，其核苷酸序列为5’-TCAGTCATGCTCTTCAGC-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-HD NI NN NG HD NI NG NN HD NG HD NG NG HD NI NN HD-C端，上述类转录激活因子(TALE)记为“TALE-5-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2)，其核苷酸序列为5’-TGCTGGTCATCCTCATC-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN HD NG NN NN NG HD NI NG HD HD NG HD NI NG HD-C端，上述类转录激活因子(TALE)记为“TALE-6-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域3)，其核苷酸序列为5’-TACACCTGCAGCTCTCAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI HD NI HD HD NG NN HD NI NN HD NG HD NG HD NI NG-C端，上述类转录激活因子(TALE)记为“TALE-7-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域3)的反义链，其核苷酸序列为5’-TGGAAATTCTTCCAGAAT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN NN NI NINI NG NG HD NG NG HD HD NI NN NI NI NG-C端，上述类转录激活因子(TALE)记为“TALE-7-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)，其核苷酸序列为5’-CTTCCTGCCCACCATCT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-HD NG NG HD HD NG NN HD HD HD NI HD HD NI NG HD NG-C端，上述类转录激活因子(TALE)记为“TALE-1’-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)的反义链，其核苷酸序列为5’-GCCCACAATGCCAGTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN HD HD HD NI HD NI NI NG NN HD HD NI NN NG NG-C端，上述类转录激活因子(TALE)记为“TALE-1’-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1’)，其核苷酸序列为5’-ACACCGAGGAAATGGGCTCA-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI HD NI HD HD NN NI NN NN NI NI NI NG NN NN NN HD NG HD NI-C端，上述类转录激活因子(TALE)记为“TALE-2’-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域1’)的反义链，其核苷酸序列为5’-CACGGAAACAGGGTTCCTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-HD NI HD NN NN NI NI NI HD NI NN NN NN NG NG HD HD NG NG-C端，上述类转录激活因子(TALE)记为“TALE-2’-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)，其核苷酸序列为5’-ATGACTCCATGAAGGAAC-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI NG NN NI HD NG HD HD NI NG NN NI NI NN NN NI NI HD-C端，上述类转录激活因子(TALE)记为“TALE-3’-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)的反义链，其核苷酸序列为5’-ATTGAAATTAGCATTTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI NG NG NN NI NI NI NG NG NI NN HD NI NG NG NG NG-C端，上述类转录激活因子(TALE)记为“TALE-3’-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)，其核苷酸序列为5’-CCTGCCCACCATCTACT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-HD HD NG NN HD HD HD NI HD HD NI NG HD NG NI HD NG-C端，上述类转录激活因子(TALE)记为“TALE-4’-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)的反义链，其核苷酸序列为5’-GCCCACAATGCCAGT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN HD HD HD NI HD NI NI NG NN HD HD NI NN NG-C端，上述类转录激活因子(TALE)记为“TALE-4’-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)，其核苷酸序列为5’-CTACTCCATCATCTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-HD NG NI HD NG HD HD NI NG HD NI NG HD NG NG-C端，上述类转录激活因子(TALE)记为“TALE-5’-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)的反义链，其核苷酸序列为5’-GACCAATCCATTGCCC-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NN NI HD HD NI NI NG HD HD NI NG NG NN HD HD HD-C端，上述类转录激活因子(TALE)记为“TALE-5’-R”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)，其核苷酸序列为5’-ATCCTGGTCATGGGTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI NG HD HD NG NN NN NG HD NI NG NN NN NN NG NG-C端，上述类转录激活因子(TALE)记为“TALE-6’-L”；

或者所述类转录激活因子(TALE)识别并结合的DNA片段来自活性区域2’)的反义链，其核苷酸序列为5’-ACTTGTCCGTCATGCTT-3’，所述类转录激活因子(TALE)与上述核苷酸序列单链识别并结合，其重复可变双氨基酸残基(repeat variable diresidue，RVD)序列为：N端-NI HD NG NG NN NG HD HD NN NG HD NI NG NN HD NG NG-C端，上述类转录激活因子(TALE)记为“TALE-6’-R”。