Human Molecular Genetics, 1999, Vol. 8, No. 10 1925-1938
© 1999 Oxford University Press
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Forward genetics in mammaliancells: functional approaches to gene discovery
1Lerner Research Institute,The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland,OH 44195, USA, 2Departmentof Genetics, Case Western Reserve University, 10900 Euclid Avenue,Cleveland, OH 44106, USA and 3Departmentof Genetics, University of Chicago, 900 South Ashland Avenue, Chicago,IL 60607, USA
ABSTRACT
Definitive proof of function in biological systemsrequires genetic analysis. Only when the loss of a particular proteincorresponds to the loss of a specific function can one be sure thatthe protein truly affects the function. Changing the pattern ofgene expression through random mutagenesis or by introducing expression libraries,followed by selection of mutant or variant cells and identificationof a missing or overexpressed protein, has the power to reveal orconfirm the roles of specific components of signaling pathways andto provide mutant cell lines and cDNA reagents to be used in definingdetailed mechanisms through structure–function analyses.These examples of forward genetics contrast with reverse geneticapproaches, where the function of a known gene product is exploredby knockout or replacement. Here we review a broad range of techniques thathave been used to alter gene expression randomly in mammalian cells,with examples of specific discoveries that have resulted from theseapplications of forward genetics.
FOOTNOTES
a To whomcorrespondence should be addressed. Tel: +1 216 444 3900;Fax: +1 216 444 3279; Email: starkg{at}ccf.org
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