Random RNAi libraries, methods of generating same, and screening methods utilizing same

摘要

This invention provides expression vectors for a ribonucleic acid (RNA) molecule comprising a double-stranded region of random sequence, sets and libraries of same, methods of generating same, and methods for identifying an RNA therapeutic or RNA molecule that has an ability to affect a biological parameter, for identifying a drug target for a disease or disorder of interest, and for identifying a variant of an RNA molecule that has an altered ability to affect a biological parameter of interest.

主权项

1. A set or library of recombinant expression vectors, wherein said set or library of recombinant expression vectors expresses a set or library of short hairpin ribonucleic acid (shRNA) molecules, each shRNA molecule comprises contiguously:
(a) a variable region 17-30 nucleotides long, consisting of a sequence, wherein said sequence is either (i) substantially random; or (ii) comprises a first sub-region and a second sub-region, wherein said first sub-region is substantially random and said second sub-region has a first sequence common to said set or library of shRNA molecules; (b) a non self-complementary region 3-20 nucleotides long, consisting of a second sequence common to said library; and (c) a complementary region consisting of a sequence, wherein said sequence is the reverse complement of the variable region; wherein the substantially random sequence in the variable region has minimal sequence bias, wherein the variable and complementary regions form a double-stranded region and the non self-complementary region forms a loop that connects the two strands of the double-stranded region.