| RNAi living-cell microarrays for loss-of-function screens in Drosophila melanogaster cells. | |
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MedLine Citation:
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PMID: 15782175 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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RNA interference (RNAi)-mediated loss-of-function screening in Drosophila melanogaster tissue culture cells is a powerful method for identifying the genes underlying cell biological functions and for annotating the fly genome. Here we describe the development of living-cell microarrays for screening large collections of RNAi-inducing double-stranded RNAs (dsRNAs) in Drosophila cells. The features of the microarrays consist of clusters of cells 200 mum in diameter, each with an RNAi-mediated depletion of a specific gene product. Because of the small size of the features, thousands of distinct dsRNAs can be screened on a single chip. The microarrays are suitable for quantitative and high-content cellular phenotyping and, in combination screens, for the identification of genetic suppressors, enhancers and synthetic lethal interactions. We used a prototype cell microarray with 384 different dsRNAs to identify previously unknown genes that affect cell proliferation and morphology, and, in a combination screen, that regulate dAkt/dPKB phosphorylation in the absence of dPTEN expression. |
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Authors:
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Douglas B Wheeler; Steve N Bailey; David A Guertin; Anne E Carpenter; Caitlin O Higgins; David M Sabatini |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't; Validation Studies Date: 2004-10-21 |
Journal Detail:
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Title: Nature methods Volume: 1 ISSN: 1548-7091 ISO Abbreviation: Nat. Methods Publication Date: 2004 Nov |
Date Detail:
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Created Date: 2005-05-03 Completed Date: 2005-05-24 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 101215604 Medline TA: Nat Methods Country: United States |
Other Details:
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Languages: eng Pagination: 127-32 Citation Subset: IM |
Affiliation:
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Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology Department of Biology, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cells, Cultured Drosophila Proteins / genetics, metabolism Drosophila melanogaster / genetics*, metabolism* Equipment Design Equipment Failure Analysis Gene Expression Profiling / instrumentation*, methods Gene Expression Regulation / physiology Genetic Testing / methods Microarray Analysis / instrumentation*, methods RNA / analysis, genetics* RNA Interference* |
| Chemical | |
Reg. No./Substance:
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0/Drosophila Proteins; 63231-63-0/RNA |
| Comments/Corrections | |
Comment In:
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Nat Methods. 2004 Nov;1(2):103-4
[PMID:
15782169
]
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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