Mutagenesis by retroviral insertion in chemical mutagen-generated quasi-haploid mammalian cells. (April 2007)

• Record Type: Journal Article
• Authors: Kim, Sung O.; Ha, Soon-Duck; Lee, Sangun; Stanton, Sarah; Beutler, Bruce; Han, Jiahuai
• Is Part Of: Biotechniques. Volume 42:Number 4(2007)
• Page Start: 493
• Page End: 501
• Publication Date: 2007-04
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000112390
• Languages: English
• ISSNs: 0736-6205

Abstract

Diploidy is a major obstacle to the mutagenic analysis of function in cultured mammalian cells. Here, we show that 6–8 rounds of chemical mutagenesis generates quasi-haploid cells that can be used as targets for insertional mutagenesis using a specially designed retroviral vector that permits rapid identification of disrupted genes in each cell that bears a phenotype of interest. The utility of combined chemical and insertional mutagenesis is illustrated by the identification of novel host genes that are required for macrophage sensitivity to anthrax lethal factor.