Kiger, A.A.;
Baum, B.;
Jones, S.;
Jones, M.R.;
Coulson, A.;
Echeverri, C.;
Perrimon, N.;
(2003)
A functional genomic analysis of cell morphology using RNA interference.
Journal of Biology
, 2
(4)
p. 27.
10.1186/1475-4924-2-27.
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Abstract
Background: The diversity of metazoan cell shapes is influenced by the dynamic cytoskeletal network. With the advent of RNA-interference (RNAi) technology, it is now possible to screen systematically for genes controlling specific cell-biological processes, including those required to generate distinct morphologies. Results: We adapted existing RNAi technology in Drosophila cell culture for use in high-throughput screens to enable a comprehensive genetic dissection of cell morphogenesis. To identify genes responsible for the characteristic shape of two morphologically distinct cell lines, we performed RNAi screens in each line with a set of double-stranded RNAs (dsRNAs) targeting 994 predicted cell shape regulators. Using automated fluorescence microscopy to visualize actin filaments, microtubules and DNA, we detected morphological phenotypes for 160 genes, one-third of which have not been previously characterized in vivo. Genes with similar phenotypes corresponded to known components of pathways controlling cytoskeletal organization and cell shape, leading us to propose similar functions for previously uncharacterized genes. Furthermore, we were able to uncover genes acting within a specific pathway using a co-RNAi screen to identify dsRNA suppressors of a cell shape change induced by Pten dsRNA. Conclusions: Using RNAi, we identified genes that influence cytoskeletal organization and morphology in two distinct cell types. Some genes exhibited similar RNAi phenotypes in both cell types, while others appeared to have cell-type-specific functions, in part reflecting the different mechanisms used to generate a round or a flat cell morphology.
| Type: | Article |
|---|---|
| Title: | A functional genomic analysis of cell morphology using RNA interference |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1186/1475-4924-2-27 |
| Publisher version: | http://dx.doi.org/10.1186/1475-4924-2-27 |
| Language: | English |
| Additional information: | © 2003 Kiger et al., licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL http://jbiol.com/content/2/4/27 |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10227 |
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