Nature Methods
- 3, 701 - 706 (2006)
Published online: 23 August 2006; | doi:10.1038/nmeth921
shRNA libraries and their use in cancer geneticsRené Bernards1, Thijn R Brummelkamp1, 2 & Roderick L Beijersbergen11
Division of Molecular Carcinogenesis and Centre for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. 2
Present address: Whitehead Institute, 9 Cambridge Center, Cambridge, Massachusetts 02142, USA.
Correspondence should be addressed to René Bernards r.bernards@nki.nl or Roderick L Beijersbergen r.beijersbergen@nki.nl RNA interference was originally described as a powerful tool to inhibit gene expression in model organisms. Until recently, loss-of-function genetic screens in mammalian cells were hampered by a lack of suitable tools that can be used in a high-throughput format. Here we discuss the construction of short-hairpin RNA (shRNA) vector libraries, in particular those generated at the Netherlands Cancer Institute (NKI), and their application in mammalian cancer genetics. We describe their virtues and limitations, as well as different options for screening such libraries.
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