Genetic inhibition by double-stranded RNA
AssigneesCarnegie Institution of Washington, University of Massachusetts
InventorsAndrew Fire, Stephen Kostas, Mary Montgomery, Lisa Timmons, SiQun Xu, Hiroaki Tabara, Samuel E. Driver, Craig C. Mello
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Abstract
A process is provided of introducing an RNA into a living cell to inhibit gene expression of a target gene in that cell. The process may be practiced ex vivo or in vivo. The RNA has a region with double-stranded structure. Inhibition is sequence-specific in that the nucleotide sequences of the duplex region of the RNA and of a portion of the target gene are identical. The present invention is distinguished from prior art interference in gene expression by antisense or triple-strand methods.
Claims19
A method to inhibit expression of a target gene in a cell in a multicellular animal comprising providing at least one ribonucleic acid (RNA) to the cell in an amount sufficient to inhibit the expression of the target gene, wherein the RNA comprises or forms a double-stranded structure containing a first strand consisting essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and a second strand consisting essentially of a ribonucleotide sequence which is complementary to the nucleotide sequence of the target gene, wherein the first and the second ribonucleotide sequences are separate complementary sequences that hybridize to each other to form said double-stranded structure, and the RNA comprising the double-stranded structure inhibits expression of the target gene, and wherein said animal is selected from the group consisting of worms and insects.
A method to inhibit expression of a target gene in a cell in an animal comprising:
- (a) isolating the cell from said animal;
- (b) introducing an expression construct into said cell encoding an RNA comprising a double-stranded structure containing a first strand consisting essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and a second strand consisting essentially of a ribonucleotide sequence which is complementary to the nucleotide sequence of the target gene, wherein the first and the second ribonucleotide sequences are separate sequences that hybridize to each other to form said double-stranded structure, and
- (c) subsequently introducing said cell into said animal, wherein said RNA comprising the double-stranded structure inhibits expression of the target gene in said cell in said animal.
A method to inhibit expression of a target gene in a cell in an invertebrate animal comprising providing at least one ribonucleic acid (RNA) to the cell in an amount sufficient to inhibit the expression of the target gene, wherein said RNA is provided to the cell by synthesizing said RNA in said cell, wherein the RNA comprises or forms a double-stranded structure containing a first strand consisting essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and a second strand consisting essentially of a ribonucleotide sequence which is complementary to the nucleotide sequence of the target gene, wherein the first and the second ribonucleotide sequences are complementary sequences that hybridize to each other to form said double-stranded structure and the RNA comprising the double-stranded structure inhibits expression of the target gene, and wherein the RNA is a double-stranded molecule containing two separate complementary RNA strands.
A method to inhibit expression of a target gene in a cell comprising:
- (a) isolating the cell from an animal;
- (b) contacting the cell with at least one ribonucleic acid (RNA) comprising a double-stranded structure containing a first strand consisting essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and a second strand consisting essentially of a ribonucleotide sequence which is complementary to the nucleotide sequence of the target gene, wherein the first and the second ribonucleotide sequences are separate sequences that hybridize to each other to form said double-stranded structure, and
- (c) subsequently introducing said cell into said animal, wherein said RNA comprising the double-stranded structure inhibits expression of the target gene in said cell.
Dependencies are inferred from the claim text.