Genetic inhibition by double-stranded RNA
View Patent ↗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.
1. A method to inhibit expression of a target gene in a cell in vitro comprising synthesizing at least two ribonucleic acids (RNAs) in the cell in an amount sufficient to inhibit the expression of a target gene, wherein the at least two RNAs form a double-stranded structure containing separate complementary strands, wherein the first RNA consists essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and the second RNA consists 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 wherein the target gene is an endogenous gene.
2. The method of claim 1 in which the cell is from an animal.
3. The method of claim 1 in which the cell is from a plant.
4. The method of claim 2 in which the cell is from an invertebrate animal.
5. The method of claim 4 in which the cell is from a nematode.
6. A method to inhibit expression of a target gene in a cell in vitro comprising synthesizing at least two ribonucleic acids (RNAs) in the cell in an amount sufficient to inhibit the expression of a target gene, wherein the at least two RNAs form a double-stranded structure containing separate complementary strands, wherein the first RNA consists essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and the second RNA consists 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 wherein the target gene is a transgene.
7. The method of claim 6 in which the cell is from an animal.
8. The method of claim 6 in which the cell is from a plant.
9. The method of claim 7 in which the cell is from an invertebrate animal.
10. The method of claim 9 in which the cell is from a nematode.
11. A method to inhibit expression of a target gene in a cell in vitro comprising synthesizing at least two ribonucleic acids (RNAs) in the cell in an amount sufficient to inhibit the expression of a target gene, wherein the at least two RNAs form a double-stranded structure containing separate complementary strands, wherein the first RNA consists essentially of a ribonucleotide sequence which corresponds to a nucleotide sequence of the target gene and the second RNA consists 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 wherein the target gene is a viral gene.
12. The method of claim 11 in which the cell is from an animal.
13. The method of claim 11 in which the cell is from a plant.
14. The method of claim 12 in which the cell is from an invertebrate animal.
15. The method of claim 14 in which the cell is from a nematode.