Mutagenesis by retroviral insertion in chemical mutagen-generated quasi-haploid mammalian cells

Sung O. Kim, Soon Duck Ha, Sangun Lee, Sarah Stanton, Bruce Beutler, Jiahuai Han

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)493-500
Number of pages8
JournalBioTechniques
Volume42
Issue number4
DOIs
StatePublished - Apr 2007

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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