High-throughput RNAi screening in cultured cells: a user's guide.
RNA interference has re-energized the field of functional genomics by enabling genome-scale loss-of-function screens in cultured cells. Looking back on the lessons that have been learned from the first wave of technology developments and applications in this exciting field, we provide both a user's guide for newcomers to the field and a detailed examination of some more complex issues, particularly concerning optimization and quality control, for more advanced users. From a discussion of cell lines, screening paradigms, reagent types and read-out methodologies, we explore in particular the complexities of designing optimal controls and normalization strategies for these challenging but extremely powerful studies.
Echeverri
Christophe J
CJ
Cenix BioScience GmbH, Tatzberg 47, Dresden 01307, Germany. echeverri@cenix-bioscience.com
Perrimon
Norbert
N
D000818:Animals; D001331:Automation; D002478:Cells, Cultured; D004331:Drosophila melanogaster / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D018014:Gene Transfer Techniques; D006801:Humans; D008954:Models, Biological; D010641:Phenotype; D011786:Quality Control; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
D034741:RNA, Small Interfering