Phenotype based functional gene screening using retrovirus-mediated gene trapping in quasi-haploid RAW 264.7 cells.
In vitro random mutagenesis, followed by phenotype screening, provides a rapid and convenient tool for identifying novel genes involved in the phenotype of interest. However, the forward mutagenic approach in mammalian somatic cells is seriously limited by the diploidic nature of the genome. To overcome this impediment, we developed a method that allows functional screening for both haploid insufficient and sufficient genes involved in the phenotype of interest, utilizing a retrovirus gene trap mutagenesis in chemical mutagen-generated quasi-haploid cells. This method was used to identify novel host genes that are required for macrophage sensitivity to anthrax lethal toxin.
Kim
Sung Ouk
SO
Infectious Diseases Research Group, Department of Microbiology and Immunology, Siebens-Drake Research Institute, University of Western Ontario, London, ON, Canada. sung.kim@schulich.uwo.ca
Ha
Soon-Duck
SD
D000818:Animals; D002460:Cell Line; D006238:Haploidy; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D012190:Retroviridae / Q000235:genetics*