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    • 7. 发明授权
    • Inhibition of PCSK9 through RNAi
    • 通过RNAi抑制PCSK9
    • US09493774B2
    • 2016-11-15
    • US13143275
    • 2010-01-05
    • Joanne KamensAnastasia Khvorova
    • Joanne KamensAnastasia Khvorova
    • C12N15/85C07H21/02C07H21/04C12N15/113
    • C12N15/1137C12N2310/14C12N2310/315C12N2310/321C12N2310/322C12N2310/344C12N2310/3515C12N2310/531C12N2320/11C12Y304/21061C12Y304/21112C12N2310/3521C12N2310/3533
    • The invention relates to various PCSK9 RNAi constructs with gene silencing activities, and uses thereof. The construct has a double-stranded region of 19-49 nucleotides, preferably 25, 26, or 27 nucleotides, and preferably blunt-ended. The construct has selective minimal modifications to confer an optimal balance of biological activity, toxicity, stability, and target gene specificity. The sense strand may be modified such that the construct is not cleaved by Dicer or other RNAse III, and the entire length of the antisense strand is loaded into RISC. In addition, the antisense strand may also be modified by 2′-O-methyl groups at the 2nd 5′-end nucleotide to greatly reduce off-target silencing. The constructs of the invention largely avoid the interferon response and sequence-independent apoptosis in mammalian cells, exhibits better serum stability, and enhanced target specificity.
    • 本发明涉及具有基因沉默活性的各种PCSK9 RNAi构建体及其应用。 该构建体具有19-49个核苷酸,优选25,26或27个核苷酸的双链区域,并且优选是平端的。 该构建体具有选择性的最小修饰以赋予生物活性,毒性,稳定性和靶基因特异性的最佳平衡。 可以修饰正义链,使得构建体不被切丝酶或其它RNA酶III切割,并且反义链的整个长度被加载到RISC中。 此外,反义链也可以在第二个5'-末端核苷酸被2'-O-甲基修饰,以大大减少脱靶沉默。 本发明的构建体大大避免了哺乳动物细胞中的干扰素反应和序列无关的凋亡,表现出更好的血清稳定性和增强的靶特异性。