Oligoribonucleotides and ribonucleases for cleaving RNA
DCFirst Claim
1. A method of activating a double-stranded RNA nuclease, comprising:
- (i) contacting the nuclease with a double-stranded oligomeric compound comprising a first oligonucleotide and a second oligonucleotide, wherein;
at least one of said first and said second oligonucleotides comprises at least four consecutive 2′
-hydroxyl ribonucleosides and at least one chemical modification;
said first and said second oligonucleotides are hybridized to each other;
said first and said second oligonucleotides are not covalently linked to each other; and
whereinsaid first and said second oligonucleotides are each independently from 15 to 25 nucleoside subunits in length; and
(ii) detecting activation of said double-stranded RNA nuclease.
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Abstract
Oligomeric compounds including oligoribonucleotides and oligoribonucleosides are provided that have subsequences of 2′-pentoribofuranosyl nucleosides that activate dsRNase. The oligoribonucleotides and oligoribonucleosides can include substituent groups for increasing binding affinity to complementary nucleic acid strand as well as substituent groups for increasing nuclease resistance. The oligomeric compounds are useful for diagnostics and other research purposes, for modulating the expression of a protein in organisms, and for the diagnosis, detection and treatment of other conditions susceptible to oligonucleotide therapeutics. Also included in the invention are mammalian ribonucleases, i.e., enzymes that degrade RNA, and substrates for such ribonucleases. Such a ribonuclease is referred to herein as a dsRNase, wherein “ds” indicates the RNase'"'"'s specificity for certain double-stranded RNA substrates. The artificial substrates for the dsRNases described herein are useful in preparing affinity matrices for purifying mammalian ribonuclease as well as non-degradative RNA-binding proteins.
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Citations
75 Claims
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1. A method of activating a double-stranded RNA nuclease, comprising:
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(i) contacting the nuclease with a double-stranded oligomeric compound comprising a first oligonucleotide and a second oligonucleotide, wherein; at least one of said first and said second oligonucleotides comprises at least four consecutive 2′
-hydroxyl ribonucleosides and at least one chemical modification;said first and said second oligonucleotides are hybridized to each other; said first and said second oligonucleotides are not covalently linked to each other; and
whereinsaid first and said second oligonucleotides are each independently from 15 to 25 nucleoside subunits in length; and (ii) detecting activation of said double-stranded RNA nuclease. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method of activating a double-stranded RNA nuclease comprising contacting the double-stranded RNA nuclease with a double-stranded oligomeric compound comprising a first oligonucleotide and a second oligonucleotide, wherein:
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said first and said second oligonucleotides are each independently 15 to 25 nucleoside subunits in length; said first and said second oligonucleotides are hybridized to each other; said first and said second oligonucleotides are not covalently linked to each other; and at least one of said first and said second oligonucleotides comprises at least four consecutive 2′
-hydroxyl ribonucleosides and at least one chemical modification. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)
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36. A method of activating a double-stranded RNA nuclease comprising contacting the double-stranded RNA nuclease with a double-stranded oligomeric compound comprising a first oligonucleotide and a second oligonucleotide, wherein:
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said first and said second oligonucleotides are each independently 15 to 25 nucleoside subunits in length; said first and said second oligonucleotides are hybridized to each other; said first and said second oligonucleotides are not covalently linked to each other; and at least one of said first and said second oligonucleotides comprises;
a plurality of nucleoside subunits with 2′
-hydroxyl pentofuranosyl sugar moieties; and
at least one chemical modification. - View Dependent Claims (37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 75)
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53. A method of activating a double-stranded RNA nuclease comprising contacting the double-stranded RNA nuclease with a double-stranded oligomeric compound comprising a first oligonucleotide and a second oligonucleotide, wherein:
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said first and said second oligonucleotides are hybridized to each other; said first and said second oligonucleotides are not covalently linked to each other; said first and said second oligonucleotides are each independently from 15 to 25 nucleoside subunits in length; and each of said first and said second oligonucleotides comprises at least four consecutive 2′
-hydroxyl ribonucleosides and at least one chemical modification. - View Dependent Claims (54, 55, 56, 57, 58, 59, 60, 61, 62, 63)
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64. A method of activating a double-stranded RNA nuclease comprising contacting the double-stranded RNA nuclease with a double-stranded oligomeric compound comprising a first oliganucleotide and a second oligonucleotide, wherein:
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said first and said second oligonucleotides are hybridized to each other; said first and said second oligonucleotides are not covalently linked to each other; said first and said second oligonucleotides are 100% complementary to each other; and at least one of said first and said second oligonucleotides comprises at least four consecutive 2′
-hydroxyl ribonucleosides and at least one chemical modification. - View Dependent Claims (65, 66, 67, 68, 69, 70, 71, 72, 73, 74)
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Specification