High-throughput RNAi screening in cultured cells: a user's guide

Nat Rev Genet. 2006 May;7(5):373-84. doi: 10.1038/nrg1836.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Automation
  • Cells, Cultured
  • Drosophila melanogaster / genetics
  • Gene Expression Profiling* / methods
  • Gene Transfer Techniques
  • Humans
  • Models, Biological
  • Phenotype
  • Quality Control
  • RNA Interference
  • RNA, Small Interfering / genetics*


  • RNA, Small Interfering