Structurally designed shRNAs
AssigneeCold Spring Harbor Laboratory
InventorsGregory J. Hannon, Sihem Cheloufi
Life of this patent
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Abstract
Provided is an improved design of shRNA based on structural mimics of miR-451 precursors. These miR-451 shRNA mimics are channeled through a novel small RNA biogenesis pathway, require AGO2 catalysis and are processed by Drosha but are independent of DICER processing. This miRNA pathway feeds active elements only into Agog because of its unique catalytic activity. These data demonstrate that this newly identified small RNA biogenesis pathway can be exploited in vivo to produce active molecules.
Claims19
A short hairpin RNA (shRNA) having the structure
- wherein X2 to X22 are nucleotides complementary to a sequence in a target gene, and are in a sequence other than the mature sequence of miR-451;
- Y4 to Y20 are nucleotides complementary to X2 to X18;
- and
- X1, Y1, Y2, and Y3 are nucleotides that are present,
- wherein, X1 and Y3 are not complementary.
A short hairpin RNA (shRNA) comprising
- (i) a first sequence of 22 nucleotides complementary to a sequence in a target gene, in a sequence other than the mature sequence of miR-451
- (ii) a second sequence directly following the first sequence, wherein the entire second sequence is fully complementary to the sequence of the first 18 nucleotides counted from the 5′ end of the first sequence,
- wherein the first sequence and the second sequence form the shRNA, and wherein the last 4 nucleotides of the first sequence form a loop region in the shRNA.
A short hairpin RNA (shRNA) comprising
- (i) a first sequence of 21 nucleotides complementary to a sequence in a target gene, in a sequence other than the mature sequence of miR-451
- (ii) a second sequence directly following the first sequence, wherein the entire second sequence is fully complementary to the sequence of the first 17 nucleotides counted from the 5′ end of the first sequence,
- wherein the first sequence and the second sequence form the shRNA, and wherein the last 4 nucleotides of the first sequence form a loop region in the shRNA.
Dependencies are inferred from the claim text.