Rapid creation and quantitative monitoring of high coverage shRNA libraries.
basic_science · Level V
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- Record sourced from PubMed, PMID 19448642.
- Also identified by DOI 10.1038/nmeth.1330 and PMC identifier 2783737.
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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
- Gene Library
- Polymerase Chain Reaction
- RNA, Messenger
- Sequence Analysis, RNA