Rapid creation and quantitative monitoring of high coverage shRNA libraries.

Bassik, Michael C; Lebbink, Robert Jan; Churchman, L Stirling; Ingolia, Nicholas T; Patena, Weronika; LeProust, Emily M; Schuldiner, Maya; Weissman, Jonathan S et al. · Nat Methods · 2009

basic_science · Level V

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Abstract

Short hairpin RNA libraries are limited by low efficacy of many shRNAs and by off-target effects, which give rise to false negatives and false positives, respectively. Here we present a strategy for rapidly creating expanded shRNA pools (approximately 30 shRNAs per gene) that are analyzed by deep sequencing (EXPAND). This approach enables identification of multiple effective target-specific shRNAs from a complex pool, allowing a rigorous statistical evaluation of true hits.

Medical subject headings