METHODS FOR MODULATING RNA USING PSEUDOCIRCULARIZATION OLIGONUCLEOTIDES
Aspects of the invention relate to methods for increasing gene expression in a targeted manner. In some embodiments, methods and compositions are provided that are useful for posttranscriptionally altering protein and/or RNA levels in a targeted manner. Aspects of the invention disclosed herein provide methods and compositions that are useful for protecting RNAs from degradation (e.g., exonuclease mediated degradation).
1 - 9 . (canceled)
10 . A method of increasing gene expression in a cell, the method comprising:
delivering to a cell an oligonucleotide comprising the general formula 5′-X 1 -X 2 -3′ wherein X 1 comprises 5 to 20 nucleotides that have a region of complementarity that is complementary with at least 5 contiguous nucleotides of an RNA transcript encoded by a gene in the cell, wherein the nucleotide at the 3′-end of the region of complementary of X 1 is complementary with the nucleotide at the transcription start site of the RNA transcript; and wherein X 2 comprises a region of complementarity that is complementary with at least 5 contiguous nucleotides of the RNA transcript that do not overlap the region of the RNA transcript that is complementary with the region of complementarity of X 1 .
11 . The method of claim 10 , wherein the region of complementarity of X 2 is within 100 nucleotides of a polyadenylation junction of the RNA transcript.
12 . The method of claim 11 , wherein the region of complementarity of X 2 is complementary with the RNA transcript immediately adjacent to or overlapping the polyadenylation junction of the RNA transcript.
13 . The method of claim 11 , wherein X 2 further comprises at least 2 consecutive pyrimidine nucleotides complementary with adenine nucleotides of the poly(A) tail of the RNA transcript.
14 . The method of claim 10 , wherein the RNA transcript is an mRNA, non-coding RNA, long non-coding RNA, miRNA, or snoRNA.
15 . The method of claim 10 , wherein the RNA transcript is an mRNA transcript, and wherein X 2 comprises a region of complementarity that is complementary with at least 5 contiguous nucleotides in the 3′-UTR of the transcript.
16 . The method claim 10 , wherein the RNA transcript is an mRNA and the delivery results in an increase in the level of a protein encoded by the mRNA.
17 - 28 . (canceled)
29 . The method of claim 10 , wherein the oligonucleotide is 10 to 50 nucleotide in length.
30 - 32 . (canceled)
33 . The method of claim 10 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.
34 . The method of claim 10 , wherein the oligonucleotide comprises at least one modified nucleotide.
35 . The method of claim 10 , wherein at least one nucleotide of the oligonucleotide comprises a 2′ O-methyl.
36 . The method of claim 10 , wherein the oligonucleotide comprises at least one ribonucleotide, at least one deoxyribonucleotide, at least one 2′-fluoro-deoxyribonucleotides or at least one bridged nucleotide.
37 . The method of claim 36 , wherein the bridged nucleotide is a LNA nucleotide, a cEt nucleotide or a ENA modified nucleotide.
38 . The method of claim 10 , wherein each nucleotide of the oligonucleotide is a LNA nucleotide.
39 . The oligonucleotide of claim 10 , wherein the nucleotides of the oligonucleotide comprise alternating deoxyribonucleotides and 2′-fluoro-deoxyribonucleotides, 2′-O-methyl nucleotides, or bridged nucleotides.
40 . The method of claim 10 , wherein the oligonucleotide is a mixmer.
41 - 155 . (canceled)