Method and medicament for inhibiting the expression of a given gene
View Patent ↗The invention relates to an isolated RNA that mediates RNA interference of an mRNA to which it corresponds and a method of mediating RNA interference of mRNA of a gene in a cell or organism using the isolated RNA.
1. A double stranded RNA (dsRNA) packaged in a liposome,
wherein said dsRNA consists of two separate complementary oligoribonucleotide strands comprising a first strand and a second strand that are linked by a linker,
wherein each of said strands consists of 21 nucleotides in length,
wherein one strand of said strands of said dsRNA is complementary to less than the full length of a third strand, said third strand comprising an RNA transcript of a mammalian target gene, and
wherein an amount of said third strand is capable of being reduced in response to said dsRNA being introduced into the presence of said third strand in a mammalian cell, thereby being capable of specifically inhibiting the expression of said mammalian target gene.
2. The dsRNA of claim 1 , wherein said separate oligoribonucleotide strands are chemically linked to reduce dissociation as compared to non-linked oligoribonucleotide strands.
3. The dsRNA of claim 2 , wherein said chemical linkage is formed by a covalent bond or hydrogen bound.
4. The dsRNA of claim 1 , wherein at least one of said oligoribonucleotide strands comprises at least one chemically modified nucleotide.
5. The dsRNA of claim 4 , wherein said chemically modified nucleotide is a 2′-modified nucleotide.
6. The dsRNA of claim 5 , wherein said 2′- modified nucleotide is a 2′-methyl substituted nucleotide.
7. The dsRNA of claim 5 , wherein said 2′- modified nucleotide is a 2′-amino substituted nucleotide.
8. The dsRNA of claim 4 , wherein said chemically modified nucleotide is a locked nucleotide.
9. The dsRNA of claim 1 , wherein said dsRNA is modified so as to be resistant to RNA degradation.
10. The dsRNA of claim 1 , wherein said two separate complementary strands are fully complementary to each other.
11. The dsRNA of claim 1 , wherein said dsRNA is capable of specifically inhibiting the expression of said target gene using dsRNA-mediated interference.
12. The dsRNA of claim 1 , wherein the linker comprises a covalent linker.
13. The dsRNA of claim 12 , wherein the covalent linker comprises a C18 linker.
14. The dsRNA of claim 1 , wherein the linker comprises a labile linker.
15. The dsRNA of claim 14 , wherein the labile linker comprises a disulfide bridge.
16. The dsRNA of claim 1 , wherein the linker is a covalent linker that links a 3′ end of one of the two strands to a 5′ end of another of the two strands.
17. The dsRNA of claim 16 , wherein the covalent linker is a C18 linker.
18. The dsRNA of claim 4 , wherein only one nucleotide of at least one of said oligoribonucleotide strands is a chemically modified nucleotide.
19. The dsRNA of claim 1 , wherein one of the strands of said dsRNA is capable of hybridizing to the RNA transcript to specifically inhibit expression of the target gene.
20. The dsRNA of claim 1 , wherein said second and third strands are sense strands and wherein said first strand is an antisense strand.
21. The dsRNA of claim 1 , wherein said first strand is capable of hybridizing to said third strand to reduce the amount of said third strand.
22. The dsRNA of claim 1 , wherein a 3′ end of one of said first and second strands is linked to a 5′ end of another of said first and second strands with a carbon-carbon linker.