Genetic inhibition by double-stranded RNA
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 an animal cell comprising introduction of a ribonucleic acid (RNA) into the cell in an amount sufficient to inhibit expression of the target gene, wherein the RNA comprises a double-stranded structure with an identical nucleotide sequence as compared to a portion of the target gene.
2. The method of claim 1 in which the target gene is a cellular gene.
3. The method of claim 1 in which the target gene is an endogenous gene.
4. The method of claim 1 in which the target gene is a transgene.
5. The method of claim 1 in which the target gene is a viral gene.
6. The method of claim 1 in which the cell is from an invertebrate animal.
7. The method of claim 6 in which the cell is from a nematode.
8. The method of claim 1 in which the identical nucleotide sequence is at least 50 bases in length.
9. The method of claim 1 in which the target gene expression is inhibited by at least 10%.
10. The method of claim 1 in which the cell is present in an organism and inhibition of target gene expression demonstrates a loss-of function phenotype.
11. The method of claim 1 in which the RNA comprises one strand which is self complementary.
12. The method of claim 1 in which the RNA comprises two separate complementary strands.
13. The method of claim 12 further comprising synthesis of the two complementary strands and initiation of RNA duplex formation outside the cell.
14. The method of claim 12 further comprising synthesis of the two complementary strands and initiation of RNA duplex formation inside the cell.
15. The method of claim 1 in which the cell is present in an organism, and the RNA is introduced within a body cavity of the organism and outside the cell.
16. The method of claim 1 in which the cell is present in an organism and the RNA is introduced by extracellular injection into the organism.
17. The method of claim 1 in which the cell is present in a first organism, and the RNA is introduced to the first organism by feeding a second, RNA-containing organism to the first organism.
18. The method of claim 17 in which the second organism is engineered to produce an RNA duplex.
19. The method of claim 1 in which an expression construct in the cell produces the RNA.