Document Detail


RNAi in Drosophila S2 cells as a tool for studying cell cycle progression.
MedLine Citation:
PMID:  19475381     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Genetic studies on model organisms, particularly yeasts and Drosophila melanogaster, have proven powerful in identifying the cell cycle machinery and its regulatory mechanisms. In more recent years RNAi has been used in a variety of genome-wide screens and single molecule studies to elucidate the mechanisms of cell cycle progression. In Drosophila cultured cells, RNAi is extremely simple, and a strong effect can be observed by adding the dsRNA to the cultured cells, with few complications of off-target effects. Functions in cell cycle progression can be followed by a variety of assays. One of the advantages of these cells is that they allow high-resolution spatiotemporal observations to be made by microscopy, with no particular complexity in terms of media and temperature. Here we discuss protocols for RNAi in Drosophila S2 culture cells, followed by the study of mitotic progression, through immunocytochemistry, live imaging, and flow cytometry analysis.
Authors:
Mónica Bettencourt-Dias; Gohta Goshima
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  545     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-05-28     Completed Date:  2009-11-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  39-62     Citation Subset:  IM    
Affiliation:
Cell Cycle Regulation Lab, Instituto Gulbenkian de Ciência, Oeiras, Portugal.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle / physiology*
Cell Line
Drosophila / cytology*
Flow Cytometry
Immunohistochemistry
Mitosis / physiology
RNA Interference*

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