Structurally designed shRNAs

摘要

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 Ago2 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.

主权项

1. A short hairpin RNA molecule (shRNA) comprising
a) (i) a first sequence of 19, 20 or 21 nucleotides fully complementary to a sequence in a target gene, having 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 15 or 16 nucleotides counted from the 5′ end of the first sequence,wherein the first sequence and the second sequence form the shRNA; or b) (i) a first sequence of 21, 22 or 23 nucleotides fully complementary to a sequence in the coding region of a target gene,
(ii) a second sequence directly following the first sequence, wherein the entire second sequence is fully complementary to the sequence of the first 17 or 18 nucleotides counted from the 5′ end of the first sequence,wherein the first sequence and the second sequence form the shRNA.