Hepatitis C dsRNA effector molecules, expression constructs, compositions, and methods of use
View Patent ↗The present invention provides agents, compositions, constructs and methods for silencing HCV polynucleotides, as well as methods and compositions for treating or preventing HCV infection in a mammalian cell. In one aspect, the present invention provides an agent or composition comprising at least one double-stranded RNA effector molecule or complex. The double-stranded RNA effector molecule or complex comprises: (1) a sequence of at least 19 nucleotides having at least 90% identity with a nucleotide sequence within HCV Conserved Region 1 (SEQ ID NO: 2), HCV Conserved Region 2 (SEQ ID NO: 3), HCV Conserved Region 5 (SEQ ID NO: 4), (ATR)-1 (SEQ ID NO: 86), ATR-2 (SEQ ID NO: 87), ATR-3 (SEQ ID NO: 88), ATR-4 (SEQ ID NO: 89); and (2) its complementary sequence. In another aspect, the present invention provides a construct suitable for replication in a host cell, and/or suitable for expression of an RNA molecule or complex of the invention in vitro or in vivo. In a third aspect, the present invention provides a method for silencing HCV RNA in a mammalian cell, which comprises administering to the mammalian cell an agent, composition, or construct of the invention in a manner and amount effective to silence HCV RNA in the cell. In a related aspect, the invention provides a method for treating or preventing HCV infection in a patient, comprising administering to the patient an effective amount of an agent, composition, or construct of the invention as described herein.
1. A method for silencing an hepatitis C virus (HCV) RNA in a mammalian cell comprising administering to said cell at least four double-stranded RNA effector molecules or complexes, comprising: (a) (1) a sequences of at least 19 consecutive nucleotides having at least 90% identity with a nucleotide sequence of SEQ ID NO: 19, SEQ ID NO: 32, SEQ ID NO: 22, and SEQ ID NO: 33; and (2) their complementary sequences; or (b) (1) sequences of at least 19 nucleotides having at least 90% identity with a nucleotide sequence of SEQ ID NO: 19, SEQ ID NO: 11, SEQ ID NO: 22, and SEQ ID NO: 33; and (2) their complementary sequences.
2. The method of claim 1 , wherein the double-stranded RNA effector molecules or complexes comprise a double-stranded region of from 19 to 30 base pairs.
3. The method of claim 2 , wherein the sequence of at least 19 nucleotides and its complementary sequence are connected via a loop sequence.
4. The method of claim 1 , wherein said administering is accomplished by providing to the mammalian cell at least one expression construct capable of expressing the double-stranded RNA effector molecule(s).
5. The method of claim 1 , wherein the mammalian cell is a human cell.
6. A method of treating hepatitis C virus (HCV) infection in a patient, comprising administering to said patient an effective amount of an agent for silencing HCV RNA, comprising at least four double stranded RNA effector molecules or complexes that comprise, or a construct that expresses: (a) (1) sequences of at least 19 nucleotides having at least 90% identity with a nucleotide sequence of SEQ ID NO: 19, SEQ ID NO: 31, SEQ ID NO: 22, and SEQ ID NO: 33; and (2) their complementary sequences; or (b) (1) sequences of at least 19 nucleotides having at least 90% identity with a nucleotide sequence of SEQ ID NO: 19, SEQ ID NO: 11, SEQ ID NO: 22, and SEQ ID NO: 33; and (2) their complementary sequences.
7. The method of claim 6 , wherein at least one double-stranded RNA effector molecule has a sequence selected from SEQ ID NOs: 50, 58, 61, 71, and 72.