Human Molecular Genetics, 1999, Vol. 8, No. 10 1925-1938
© 1999 Oxford University Press
Review |
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
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  T. Lu, M. W. Jackson, A. D. Singhi, E. S. Kandel, M. Yang, Y. Zhang, A. V. Gudkov, and G. R. Stark Validation-based insertional mutagenesis identifies lysine demethylase FBXL11 as a negative regulator of NF{kappa}B PNAS, September 22, 2009; 106(38): 16339 - 16344. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. A. Newman, S. J. Muh, R. H. Hovhannisyan, C. C. Warzecha, R. B. Jones, W. L. McKeehan, and R. P. Carstens Identification of RNA-binding proteins that regulate FGFR2 splicing through the use of sensitive and specific dual color fluorescence minigene assays RNA, June 1, 2006; 12(6): 1129 - 1141. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Pritsker, N. R. Ford, H. T. Jenq, and I. R. Lemischka Genomewide gain-of-function genetic screen identifies functionally active genes in mouse embryonic stem cells PNAS, May 2, 2006; 103(18): 6946 - 6951. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. S. Kandel, T. Lu, Y. Wan, M. K. Agarwal, M. W. Jackson, and G. R. Stark Mutagenesis by reversible promoter insertion to study the activation of NF-{kappa}B PNAS, May 3, 2005; 102(18): 6425 - 6430. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. R. Stark My Life in Science, Not the Restaurant Business J. Biol. Chem., March 18, 2005; 280(11): 9753 - 9760. [Full Text] [PDF]
|
|
|
|
|
|
A. D. Singhi, R. V. Kondratov, N. Neznanov, M. V. Chernov, and A. V. Gudkov Selection-subtraction approach (SSA): A universal genetic screening technique that enables negative selection PNAS, June 22, 2004; 101(25): 9327 - 9332. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Sharifi Gene Function Far From Understood Mayo Clin. Proc., December 1, 2002; 77(12): 1395 - 1396. [PDF]
|
|
|
|
|
|
X. Li, M. Commane, H. Nie, X. Hua, M. Chatterjee-Kishore, D. Wald, M. Haag, and G. R. Stark Act1, an NF-kappa B-activating protein PNAS, August 23, 2000; (2000) 160265197. [Abstract] [Full Text]
|
|
|
|
 |
|
 J. Gécz and J. Mulley Genes for Cognitive Function: Developments on the X Genome Res., February 1, 2000; 10(2): 157 - 163. [Abstract] [Full Text]
|
|
|
|
|
|
X. Li, M. Commane, H. Nie, X. Hua, M. Chatterjee-Kishore, D. Wald, M. Haag, and G. R. Stark Act1, an NF-kappa B-activating protein PNAS, September 12, 2000; 97(19): 10489 - 10493. [Abstract] [Full Text] [PDF]
|
|