4.8 Article

Rapid creation and quantitative monitoring of high coverage shRNA libraries

Journal

NATURE METHODS
Volume 6, Issue 6, Pages 443-U62

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1330

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Funding

  1. Netherlands Organization for Scientific Research (NWO)
  2. Leukemia and Lymphoma Society
  3. US National Institutes of Health [K99/R00, R01 GM80783]
  4. National Institute of General Medical Sciences [GM080853]
  5. Sandler New Technologies
  6. Fight For Mike foundation

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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 (similar to 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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