Antigene oligomers inhibit transcription

Abstract

Transcription of a gene in a mammalian cell is methylase-independently inhibited by contacting the cell with a nucleic acid oligomer of 12-28 bases complementary to a partially single-stranded target genomic sequence of the gene.

1 . A method of methylase-independently inhibiting transcription of a gene in a mammalian cell, the method comprising the steps of:
 contacting the cell with a nucleic acid oligomer of 12-28 bases complementary to a partially single-stranded target genomic sequence of the gene, whereby the transcription of the gene is subject to a resultant methylase-independent inhibition; and   detecting the resultant methylase-independent inhibition.   
     
     
         2 . The method of  claim 1  wherein the target genomic sequence is selected from the group consisting of an AT-rich sequence, a cruciform loop, a G-quadruplex, a nuclease hypersensitive element (NHE), and an open complex region located between nucleotides −50 to +25 from the gene's transcription start site. 
     
     
         3 - 5 . (canceled) 
     
     
         6 . The method of  claim 1  wherein the target genomic sequence includes the transcription start site. 
     
     
         7 . The method of  claim 1  wherein the oligomer is selected from the group consisting of a double-stranded RNA, a DNA, a peptide nucleic acid, and a morpholino. 
     
     
         8 . The method of  claim 1  wherein the oligomer is a double-stranded RNA, and the target genomic sequence is not within a CpG island, and/or does not include a CG dinucleotide. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 1  wherein the oligomer is a double-stranded RNA of 18-25 bases. 
     
     
         11 . The method of  claim 1 , wherein the oligomer is double-stranded RNA comprising a nucleotide having a 2′ chemical modification. 
     
     
         12 . The method of  claim 1 , wherein the oligomer is double-stranded RNA comprising a serum stability-enhancing chemical modification selected from the group consisting of a phosphorothioate internucleotide linkage, a 2′-O-methyl ribonucleotide, a 2′-deoxy-2′-fluoro ribonucleotide, a 2′-deoxy ribonucleotide, a universal base nucleotide, a 5-C-methyl nucleotide, an inverted deoxyabasic residue incorporation, and a locked nucleic acid. 
     
     
         13 . The method of  claim 1  wherein the cell is a cultured cell in vitro. 
     
     
         14 . The method of  claim 1  wherein the cell is in situ in a host. 
     
     
         15 . The method of  claim 1  wherein the cell is a cultured cell in vitro, the oligomer is a double-stranded RNA of 18-25 bases, and the cell is contacted with a composition comprising the oligomer and a cationic lipid. 
     
     
         16 . The method of  claim 1  wherein the cell is in situ in a host, the oligomer is a double-stranded RNA of 18-25 bases, and the cell is contacted with a composition consisting essentially of the oligomer. 
     
     
         17 . The method of  claim 1  wherein the contacting step is free of viral transduction. 
     
     
         18 . The method of  claim 17  wherein the cell is contacted with a composition consisting essentially of the oligomer. 
     
     
         19 . The method of  claim 17  wherein the inhibition is at least 90%. 
     
     
         20 . The method of  claim 1  wherein the oligomer is a double-stranded RNA of 18-25 bases, the inhibition is at least 90%, and a single sequence of the gene is targeted. 
     
     
         21 . The method of  claim 17  wherein the oligomer is not attached to a nuclear localization peptide. 
     
     
         22 . The method of  claim 1  wherein the cell is contacted with a 1-100 nanomolar concentration of the oligomer. 
     
     
         23 . The method of  claim 1  wherein the cell is a breast carcinoma cell and the gene encodes the human progesterone receptor, or wherein the cell is a breast carcinoma cell and the gene encodes the human androgen receptor, or wherein the cell is a retina cell and the gene encodes the vascular endothelial growth factor (VEGF). 
     
     
         24 - 25 . (canceled) 
     
     
         26 . A nucleic acid oligomer for methylase-independent inhibition of transcription of a human progesterone receptor gene, a human androgen receptor gene, or a human vascular endothelial growth factor gene, the oligomer comprising a nucleotide sequence of 12-28 bases complementary to a target sequence located between nucleotides −50 to +25 of the transcription start site of the gene. 
     
     
         27 - 35 . (canceled)