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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2783737 | PMC |
| http://dx.doi.org/10.1038/nmeth.1330 | DOI Listing |
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