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US 8,202,846 B2Granted
Methods and compositions for RNA interference
AssigneeCold Spring Harbor Laboratory
InventorsGregory J. Hannon, Patrick Paddison, Emily Bernstein, Amy Caudy, Douglas Conklin, Scott Hammond
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Abstract
The present invention provides methods for attenuating gene expression in a cell, especially in a mammalian cell, using gene-targeted double stranded RNA (dsRNA), such as a hairpin RNA. The dsRNA contains a nucleotide sequence that hybridizes under physiologic conditions of the cell to the nucleotide sequence of at least a portion of the gene to be inhibited (the “target” gene).
Claims10
A method for attenuating expression of a target gene in a mammalian cell, the method comprising:
- introducing into a mammalian cell an expression vector comprising:
- (i) an RNA polymerase promoter, and
- (ii) a sequence encoding a short hairpin RNA molecule comprising a double-stranded region, wherein the double-stranded region consists of at least 20 nucleotides but not more than 29 nucleotides,
- wherein the short hairpin RNA molecule is a substrate for Dicer-dependent cleavage and does not trigger a protein kinase RNA-activated (PK) response in the mammalian cell,
- wherein the double-stranded region of the short hairpin RNA molecule comprises a sequence that is complementary to a portion of the target gene, and
- wherein the short hairpin RNA molecule is stably expressed in the mammalian cell in an amount sufficient to attenuate expression of the target gene in a sequence specific manner, and is expressed in the cell without use of a PK inhibitor, whereby expression of the target gene is inhibited.
3 more elements
Dependencies are inferred from the claim text.