Cre-lox based gene knockdown constructs and methods of use thereof

摘要

The present invention relates to vectors, compositions and methods for conditional, Cre-lox regulated, RNA interference. The vectors allow for spatial and temporal control of miRNA expression in vivo.

主权项

1. A vector comprising
a. a first pair of loxP sequences inverted in orientation with respect to each other, b. a nucleic acid cassette comprising a first nucleic acid encoding
(i) at least one nucleotide sequence that can form a hairpin structure comprising at least one miRNA agent, and(ii) miRNA sequences flanking the at least one nucleotide sequence, wherein the miRNA sequences flanking the at least one nucleotide sequence are derived from endogenous miRNA sequences that flank wild-type hairpin structures, and wherein the nucleic acid cassette is positioned in an antisense orientation between the first pair of loxP sequences, c. a second pair of loxP sequences distinct from the first pair of the loxP sequences, inverted in orientation with respect to each other, wherein the first pair of loxP sequences and the second pair of loxP sequences are configured such that flipping of the first pair results in excision of the sequences between the second pair.