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. An oligoribonucleotide of double-stranded structure (dsRNA) for inhibiting the expression of a given target gene in mammalian cells, wherein at least one strand of the dsRNA comprises a locked nucleotide, wherein at least one strand of the dsRNA comprises at least one chemically modified nucleotide that is distinct from the locked nucleotide, and wherein the dsRNA has 15 to 49 base pairs and has a complementary region I that is incorporated in the dsRNA.
2. The dsRNA of claim 1 , wherein the complementary region I has not more than 49 nucleotide pairs.
3. The dsRNA of claim 1 , wherein the chemically modified nucleotide is a 2′-modified nucleotide.
4. The dsRNA of claim 3 , wherein the 2′-modified nucleotide is a 2′-methyl substituted nucleotide.
5. The dsRNA of claim 3 , wherein the 2′-modified nucleotide is a 2′-amino substituted nucleotide.
6. The dsRNA of claim 1 , wherein the locked nucleotide is chemically modified by a 2′-O,4′-C methylene bridge.
7. The dsRNA of claim 1 , wherein the target gene is a mammalian gene or a viral gene.
8. The dsRNA of claim 1 , wherein the target gene is selected from the group consisting of an oncogene, a cytokine gene, an Id protein gene, a developmental gene, a PKR gene and a prion gene.
9. The dsRNA of claim 1 , wherein the dsRNA is enclosed by a micellar structure.
10. The dsRNA of claim 9 , wherein the micellar structure comprises a liposome.
11. The dsRNA of claim 1 , wherein the target gene is expressed in eukaryotic cells.
12. The dsRNA of claim 1 , wherein an end of the dsRNA is modified in order for the double-stranded structure to counteract degradation.
13. The dsRNA of claim 1 , wherein the dsRNA comprises a sense strand and an antisense strand, and wherein the sense strand comprises a 2′-methyl substituted nucleotide.
14. The dsRNA of claim 1 , wherein the dsRNA comprises a sense strand and an antisense strand, and wherein the sense strand comprises a 2′-methoxynucleotide.
15. The dsRNA of claim 1 , wherein the dsRNA comprises a sense strand and an antisense strand, and wherein the sense strand comprises a plurality of 2′-methoxynucleotides.
16. The dsRNA of claim 1 , wherein the dsRNA comprises a pharmaceutical composition formulated for delivery to a mammalian cell.
17. A pharmaceutical composition comprising an oligoribonucleotide of double-stranded structure (dsRNA) for inhibiting the expression of a given target gene in mammalian cells, wherein at least one strand of the dsRNA comprises a locked nucleotide, wherein at least one strand of the dsRNA comprises at least one chemically modified nucleotide that is distinct from the locked nucleotide, and wherein the dsRNA has 15 to 49 base pairs and has a complementary region I that is incorporated in the dsRNA.
18. The pharmaceutical composition of claim 17 , further comprising a liposome enclosing the dsRNA for delivery to a mammalian cell.