Gene trapping in embryonic stem cells.
Gene trapping in embryonic stem cells (ESCs) generates random, sequence-tagged insertional mutations, which can often report the gene expression pattern of the mutated gene. This mutagenesis strategy has often been coupled to expression or function-based assays in gene discovery screens. The availability of the mouse genome sequence has shifted gene trapping from a gene discovery platform to a high-throughput mutagenesis platform. At present, a concerted worldwide effort is underway to develop a library of loss-of-function mutations in all mouse genes. The International Gene Trap Consortium (IGTC) is leading the way by making a first pass of the genome by random mutagenesis before a high-throughput gene targeting program takes over. In this chapter, we provide a methods guidebook to exploring and using the IGTC resource, explain the different kinds of vectors and insertions that reside in the different libraries, and provide advice and methods for investigators to design novel expression-based "cottage industry" screens.
Stanford
William L
WL
Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
Epp
Trevor
T
Reid
Tammy
T
Rossant
Janet
J
D000818:Animals; D018929:Cell Culture Techniques; D002999:Clone Cells; D019295:Computational Biology; D053595:Embryonic Stem Cells / Q000145:classification / Q000166:cytology / Q000378:metabolism; D020224:Expressed Sequence Tags / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D016254:Mutagenesis, Insertional