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    • 9. 发明申请
    • Composition and method for introduction of RNA interference sequences into targeted cells and tissues
    • 将RNA干扰序列引入目标细胞和组织的组合物和方法
    • US20050260214A1
    • 2005-11-24
    • US11126551
    • 2005-05-11
    • Michael Simon
    • Michael Simon
    • A61K39/395A61K47/48A61K48/00C07K16/28C07K16/46
    • C07K16/2896A61K47/6807A61K47/6849C07K2317/54
    • A composition and method are provided by which double-stranded RNA containing small interfering RNA nucleotide sequences is introduced into specific cells and tissues for the purpose of inhibiting gene expression and protein production in those cells and tissues. Intracellular introduction of the small interfering RNA nucleotide sequences is accomplished by the internalization of a target cell specific ligand to which the double-stranded RNA containing a small interfering RNA nucleotide sequence is conjugated. The ligand is specific to a unique target cell surface antigen. The ligand is either spontaneously internalized after binding to the cell surface antigen. Internalization is also facilitated by the binding of an RNA binding protein to the double-stranded RNA. If the unique cell surface antigen is not naturally internalized after binding to its ligand, internalization is promoted by the incorporation of an arginine-rich peptide, or other membrane permeable peptide, into the structure of the ligand or attachment of such a peptide to the ligand. The composition and method are practiced in whole living mammals, as well as cells living in tissue culture. The dsRNA is then hydrolyzed by Dicer, an RNAse III-like ribonuclease, thereby releasing siRNA which silences the target gene. Inhibition is nucleotide sequence specific and depends upon sequence identity of small interfering RNA with the target nucleic acid.
    • 提供了一种组合物和方法,其中含有小干扰RNA核苷酸序列的双链RNA被引入到特定细胞和组织中,目的是抑制这些细胞和组织中的基因表达和蛋白质产生。 小干扰RNA核苷酸序列的细胞内引入是通过将含有小干扰RNA核苷酸序列的双链RNA共轭的靶细胞特异性配体内化来实现的。 该配体对独特的靶细胞表面抗原是特异性的。 配体在与细胞表面抗原结合后自发内化。 通过RNA结合蛋白与双链RNA的结合也促进了内化。 如果独特的细胞表面抗原在与其配体结合后不被天然内化,则通过将富含精氨酸的肽或其它膜可渗透肽掺入配体的结构或将这种肽连接到配体来促进内化 。 组合物和方法在整个活的哺乳动物以及生活在组织培养中的细胞中实践。 然后通过Dicer(一种RNA酶III样核糖核酸酶)水解dsRNA,从而释放沉默靶基因的siRNA。 抑制是核苷酸序列特异性,取决于小干扰RNA与靶核酸的序列同一性。