Pooled shRNA screenings: experimental approach.
RNA interference (RNAi) has emerged as a powerful genetic strategy to functionally interrogate the entire genome by loss-of-function studies. In the last years, several arrayed shRNA libraries that can silence almost all the human genome have been developed. The generation of new and more efficient shRNA plasmids has allowed performing genetic screens in a pooled fashion and provides the opportunity to investigate the entire genome finding relevant genes to any biological process. In this chapter, the pipeline and methods to perform a pooled shRNA screen are discussed.
Institute for Cancer Genetics, Columbia University, New York, NY, USA.
Ruth
R
Rodriguez-Barrueco
Netonia
N
Marshall
Jose M
JM
Silva
D000818:Animals; D005434:Flow Cytometry; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D006801:Humans; D051379:Mice; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014161:Transduction, Genetic; D014169:Transformation, Bacterial
D034741:RNA, Small Interfering