RNAi screening

RNA interference has enabled genome-scale loss-of-function screens in cultured cells. The fact that RNAi can be scaled up for use in high-throughput techniques has motivated the creation of genome-wide RNAi reagents.

Comprehensive, high-throughput analysis of gene function using RNA interference (RNAi)-based screens is emerging as a significant step forward for preliminary drug-target identification.

Until quite recently drug target identification depended heavily on the analysis of changes in gene expression, which in turn needed to be correlated with gene function.

The promise of obtaining preliminary ’gene to phenotype’ information using a single high-throughput platform is propelling major investment in this area by biotechnology and pharmaceutical companies.

High-throughput genetic screens have exponentially increased the functional annotation of the genome over the past 10 years.

Likewise, genome-scale efforts to map DNA methylation, chromatin state and occupancy, messenger RNA expression patterns, and disease-associated genetic polymorphisms, and proteome-wide efforts to map protein-protein interactions, have also created vast resources of data.

An emerging trend involves combining multiple types of data, referred to as integrative screening. Several studies demonstrate the power of data integration to generate focused, validated data sets and to identify high-confidence candidate genes for follow-up experiments. (19436058)

See also

 RNAi screening and cancer

References

 RNAi-based screening identifies the Mms22L-Nfkbil2 complex as a novel regulator of DNA replication in human cells. Piwko W, Olma MH, Held M, Bianco JN, Pedrioli PG, Hofmann K, Pasero P, Gerlich DW, Peter M. EMBO J. 2010 Dec 15;29(24):4210-22. PMID: 21113133

 Integrative analysis of genome-wide RNA interference screens.
Berndt JD, Biechele TL, Moon RT, Major MB. Sci Signal. 2009 May 12;2(70):pt4. PMID: 19436058

 Genome-wide screening for gene function using RNAi in mammalian cells. Cullen LM, Arndt GM. Immunol Cell Biol. 2005 Jun;83(3):217-23. PMID: 15877598

 Boutros M, Ahringer J. The art and design of genetic screens: RNA interference. Nat Rev Genet. 2008 Jun 3. PMID: 18521077

 Chatterjee-Kishore M. From genome to phenome—RNAi library screening and hit characterization using signaling pathway analysis. Curr Opin Drug Discov Devel. 2006 Mar;9(2):231-9.PMID: 16566293

 Moffat J, Sabatini DM. Building mammalian signalling pathways with RNAi screens. Rev Mol Cell Biol. 2006 Mar;7(3):177-87.
PMID: 16496020

 Echeverri CJ, Perrimon N. High-throughput RNAi screening in cultured cells: a user’s guide. Nat Rev Genet. 2006 May;7(5):373-84. PMID: 16607398

 Zhu C, Zhao J, Bibikova M, Leverson JD, Bossy-Wetzel E, Fan JB, Abraham RT, Jiang W. Functional analysis of human microtubule-based motor proteins, the kinesins and dyneins, in mitosis/cytokinesis using RNA interference. Mol Biol Cell. 2005 Jul;16(7):3187-99. PMID: 15843429