High potency siRNAS for reducing the expression of target genes
First Claim
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1. A method for making siRNA of increased potency comprising:
- (a) obtaining nucleotides;
(b) incorporating the nucleotides into siRNA such that an RNA duplex of from 15 to 30 contiguous nucleotides is formed, wherein the siRNA has a sequence that is substantially identical to at least a portion of a selected target gene.
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Abstract
The present invention provides improved methods of attenuating gene expression through the phenomenon of RNA interference. The invention provides methods of synthesis of double stranded RNAs (dsRNAs) of increased potency for use as small interfering RNA (siRNA). Surprisingly and unexpectedly, siRNAs made by the methods of the invention are significantly more potent than previously available siRNAs.
294 Citations
129 Claims
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1. A method for making siRNA of increased potency comprising:
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(a) obtaining nucleotides;
(b) incorporating the nucleotides into siRNA such that an RNA duplex of from 15 to 30 contiguous nucleotides is formed, wherein the siRNA has a sequence that is substantially identical to at least a portion of a selected target gene. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
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46. A method for attenuating the expression of a target gene in a cell comprising:
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(a) obtaining a siRNA of increased potency;
(b) introducing the siRNA of increased potency into the cell in an amount sufficient to attenuate expression of the target gene. - View Dependent Claims (47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99)
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- 100. A siRNA of increased potency comprising a duplex structure of from 15 to 30 nucleotides that has a nucleotide sequence substantially identical to at least a portion of a target gene.
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124. A cell comprising a target gene whose expression is attenuated by a method comprising:
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(a) obtaining siRNA of increased potency comprising a duplex structure of from 15 to 30 nucleotides that has a nucleotide sequence substantially identical to at least a portion of a target gene; and
(b) introducing the siRNA of increased potency into the cell in an amount sufficient to attenuate expression of the target gene.
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- 125. A kit for making siRNA of increased potency comprising nucleotides.
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129. A kit for attenuating expression of a target gene in a cell comprising at least one siRNA of increased potency.
Specification