Mutagenesis by retroviral insertion in chemical mutagen-generated quasi-haploid mammalian cells.
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.
Kim
Sung O
SO
Department of Microbiology, University of Western Ontario, London, ON, Canada. sung.kim@schulich.uwo.ca
Ha
Soon-Duck
SD
Lee
Sangun
S
Stanton
Sarah
S
Beutler
Bruce
B
Han
Jiahuai
J
D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D006238:Haploidy; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D009153:Mutagens; D009154:Mutation; D010641:Phenotype; D012190:Retroviridae / Q000235:genetics*
D009153:Mutagens