Compositions for RNA interference and methods of use thereof

• US 8,729,036 B2
• Filed: 08/07/2003
• Issued: 05/20/2014
• Est. Priority Date: 08/07/2002
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Zamore, Phillip D., Hutvgner, Gyrgy, Schwarz, Dianne, Tang, Guiliang, Haley, Benjamin
• Primary Examiner(s): SHIN, DANA H
• Application Number:
  US10/638,253
• Publication Number:
  US 20040203145A1
• Time in Patent Office: 3,939 Days
• Field of Search: 514/44, 435/4, 435/6, 435/375, 536/23.1, 536/24.5
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2330/50 Biochemical production, i.e. in a transformed host cell

Abstract

The present invention provides compositions for RNA interference and methods of use thereof. In particular, the invention provides single-stranded small interfering RNAs. Functional and genomic and proteomic methods are featured. Therapeutic methods are also featured.

Claims

1. A method of activating target-specific RNA interference (RNAi) in a mammalian cell in vitro, comprising introducing into said cell an isolated, single-stranded small interfering RNA molecule (ss-siRNA) having a length of 19-21 nucleotides, wherein the sequence of said ss-siRNA molecule is complementary to a target mRNA sequence and directs target-specific RNA interference (RNAi) and wherein the 5′

nucleotide is 5′

phosphorylated or is capable of being 5′

phosphorylated in vitro, said ss-siRNA being introduced in an amount sufficient for degradation of the target mRNA to occur, thereby activating target-specific RNAi in the cell.

2. The method of claim 1, wherein the ss-siRNA is introduced into the cell by contacting the cell with the ss-siRNA.

3. The method of claim 2, wherein the ss-siRNA is introduced into the cell by contacting the cell with a composition comprising the ss-siRNA and a lipophillic carrier.

4. The method of claim 1, wherein the target mRNA specifies the amino acid sequence of a protein involved or predicted to be involved in a human disease or disorder.

5. The method of claim 1, wherein degradation of the target mRNA produces a loss-of-function phenotype.

6. The method of claim 1, wherein the ss-siRNA is modified by the substitution of the 3′

-OH of the 3′

most nucleotide.

7. The method of claim 1, wherein the 3′

most nucleotide has a 3′

-OH replaced by a 2′

, 3′

dideoxy terminus or an amino modifier group.

8. The method of claim 1, wherein the ss-siRNA has a length of 19 nucleotides.

9. The method of claim 1, wherein the ss-siRNA has a length of 20 nucleotides.

10. The method of claim 1, wherein the ss-siRNA has a length of 21 nucleotides.

11. The method of claim 1, wherein the 5′

nucleotide of the ss-siRNA is 5′

phosphorylated with a phosphate groups selected from the group consisting of a momophosphate, a diphosphate and a triphosphate.