MODULATION OF LDL RECEPTOR GENE EXPRESSION WITH DOUBLE-STRANDED RNAS TARGETING THE LDL RECEPTOR GENE PROMOTER
Gene expression can be selectively regulated by double-stranded “antigene” RNAs that target regions of the low density lipoprotein receptor (LDL-R) promoter, thereby permitting modulation of LDL levels in vivo and subsequent effects on circulating LDL levels.
1 . A method of modulating expression of low density lipoprotein receptor (LDL-R) in a cell comprising contacting said cell with a first double-stranded RNA complementary to a portion of an LDL-R promoter.
2 . The method of claim 1 , wherein the double-stranded RNA increases LDL-R expression.
3 . The method of claim 1 , wherein the double-stranded RNA decreases LDL-R expression.
4 . The method of claim 1 , wherein the double-stranded RNA targets a Repeat 2 region, a Repeat 3 region or both.
5 . The method of claim 1 , wherein the double-stranded RNA targets a sterol-independent regulatory element.
6 . The method of claim 1 , wherein the double-stranded RNA is complementary a region within bases −1 to −200, relative to the transcription start site, of the LDL-R gene.
7 . (canceled)
8 . The method of claim 1 , wherein the double-stranded RNA contains one or more modified nucelosides.
9 . (canceled)
10 . (canceled)
11 . The method of claim 8 , wherein one strand of the double-stranded RNA contains one or more modified nucelosides, and the other strand does not contain a modified nuceloside.
12 . (canceled)
13 . (canceled)
14 . The method of claim 1 , wherein said cell is located in situ in a host, and the contacting step is effected by administering to the host an effective amount of the double-stranded RNA.
15 . (canceled)
16 . The method of claim 1 , further comprising detecting a change in the expression of LDL-R.
17 . The method of claim 16 , wherein detecting comprises inferring a change in the expression from a physiologic change in the cell.
18 . The method of claim 17 , wherein the cell is located in situ in a host and detecting comprises inferring a change in the expression from a physiologic change in the host.
19 . The method of claim 16 , wherein detecting comprises one or more of Northern blot, PCR, immunohistochemistry, Western blot or ELISA.
20 . The method of claim 1 , wherein the RNA further comprises one or more deoxyribonucleotides.
21 . The method of claim 20 , wherein the RNA comprises a dTdT dinucleotide overhang of each strand.
22 . The method of claim 20 , wherein the RNA comprises at least one phosphorothioate linkage in each strand.
23 .- 25 . (canceled)
26 . The method of claim 1 , wherein said double-stranded RNA is formulated in a lipid vehicle.
27 . A method of reducing circulating low density lipoprotein in a subject comprising administering to said subject a first double-stranded RNA complementary to a portion of an LDL-R promoter.
28 . The method of claim 27 , wherein the subject suffers from hypercholesterolemia, atherolsclerosis and/or coronary heart disease.
29 .- 35 . (canceled)
36 . The method of claim 27 , wherein said double-stranded RNA is formulated in a lipid vehicle.
37 . A pharmaceutical formulation comprising (a) a double-stranded RNA complementary to a portion of a low density lipoprotein receptor (LDL-R) promoter, and (b) a pharmaceutically acceptable buffer, carrier or diluent.
38 .- 43 . (canceled)