Lipid formulated compositions and methods for inhibiting expression of Serum Amyloid A gene
The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting a Serum Amyloid A (SAA) gene, and methods of using the dsRNA to inhibit expression of SAA.
1. A double-stranded ribonucleic acid (dsRNA), wherein said dsRNA comprises a sense strand consisting of the nucleotide sequence of SEQ ID NO:189 and an antisense strand consisting of the nucleotide sequence of SEQ ID NO:190.
2. The dsRNA of claim 1 , wherein the sense strand consists of SEQ ID NO:33 and the antisense strand consists of SEQ ID NO:34.
3. The dsRNA of claim 1 , comprising at least one modified nucleotide.
4. The dsRNA of claim 1 , comprising at least one modified nucleotide selected from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a 2′-O-methylcytidine-5′-phosphate, a 2′-O-methyluridine-5′-phosphate, and a non-natural base comprising nucleotide.
5. The dsRNA of claim 1 , comprising a dTdT overhang on the 3′ end of each strand.
6. The dsRNA of claim 1 , wherein the dsRNA is conjugated to a ligand.
7. The dsRNA of claim 1 , wherein the dsRNA is formulated in a lipid formulation.
8. The dsRNA of claim 1 , wherein administration of the dsRNA to a cell results in 92% inhibition of SAA mRNA expression as measured by a real time PCR assay or 85% inhibition of SAA mRNA expression as measured by a branched DNA assay or 100% inhibition of SAA protein expression as measured by a ELISA assay compared to an siRNA control.
9. The dsRNA of claim 1 , wherein the dsRNA has an IC50 of less than 10 pM as measured by a branched DNA assay.
10. A cell comprising the dsRNA of claim 1 .
11. A vector comprising a nucleotide sequence encoding at least one strand of the dsRNA of claim 1 .
12. A cell comprising the vector of claim 11 .
13. A pharmaceutical composition for inhibiting expression of an SAA gene comprising the dsRNA of claim 1 .
14. A method of inhibiting SAA expression in a cell, the method comprising:
(a) introducing into the cell the dsRNA of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an SAA gene, thereby inhibiting expression of the SAA gene in the cell.
15. The method of claim 14 , wherein the method is performed in vitro.
16. A pharmaceutical composition for inhibiting expression of an SAA gene comprising the dsRNA of claim 2 .
17. A method of inhibiting SAA expression in a cell, the method comprising:
(a) introducing into the cell the dsRNA of claim 2 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an SAA gene, thereby inhibiting expression of the SAA gene in the cell.
18. The method of claim 17 , wherein the method is performed in vitro.