International Mammalian Genome Society

17th International Mouse Genome Conference

9-12 November 2003, Braunschweig, Germany

Plenary Presentations * Oral Presentations *
Poster Presentations: Behavioural Genetics and Genomics * Development and Stem Cells * Functional Genome Analysis * Mouse Models of Human Disease * Mouse System Biology Bioinformatics * Multigenic and Multifactorial Trait Analysis * Nutrition and Metabolic Disease * Phenotyping Methods Imaging * The Genetics and Genomics of Infectious Disease *
Verne Chapman Memorial Lecture * Table of Contents * Sponsor/Exhibitor List * Awards * Photographs

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POSTER 32 - AN EXPRESSION GENE TRAP MUTAGENESIS RESOURCE AT THE CENTRE FOR MODELING HUMAN DISEASE (CMHD)

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1) Institute of Biomaterials & Biomedical Engineering, University of Toronto, Toronto, Canada, 3) Programme in Development and Fetal Health, Samuel Lunenfeld Research Institute, Toronto, Canada, 4) Centre for Modeling Human Disease, Toronto, Canada

Co-Authors: 1) 3) 4) Reid T, 4) Hant P, 1) 3) 4) Lan Q, 1) 3) 4) Li C, 1) 3) 4) Ohishi M, 4) To C, 4) Tsao N, 1) 3) 4) Yu M, 4) Kassam N, 2) 4) Osborne L, 2) 3) 4) Rossant J, 1) 3) 4) Stanford W
Institutions: 1) Institute of Biomaterials & Biomedical Engineering, University of Toronto, Toronto, Canada, 2) Department of Medical Genetics, University of Toronto, Toronto, Canada, 3) Programme in Development and Fetal Health, Samuel Lunenfeld Research Institute, Toronto, Canada, 4) Centre for Modeling Human Disease, Toronto, Canada

Gene trap mutagenesis of mouse embryonic stem (ES) cells generates random loss-of-function mutations, which can be identified by a sequence tag and can often report the endogenous expression of the mutated gene. The Centre for Modeling Human Disease (CMHD) is performing gene trap-based expression and genotypic screens to generate new mouse mutations as a resource for the scientific community. The gene trap insertions are screened using multiplexed in vitro differentiation and induction assays. Data is stored in a MySQL relational database and a user-friendly web-based interface allows researchers to make advanced queries of the expression data. Sequence tags are being generated by 5'-RACE, 3'-RACE, and inverse PCR to complement expression profiles. Semi-automated sequence processing and annotation algorithms are used to identify trapped genes and to extract additional information including chromosomal location, gene ontology (GO) classifications, and protein functional domains. The sequence tag database can currently be queried using a BLAST interface and, by the time of this meeting researchers will also be able to make advanced queries of sequence annotations using Boolean operators. We will discuss the randomness of genomic integration and intragenic insertion of traditional gene trap vectors with more recent innovations in gene trap vector design, such as the incorporation of a 3' splice donor sequence designed to make transcription of the selectable marker contingent upon trapping a polyadenylation site. We have also begun to generate mice from polyA trapped clones to address the mutagenicity of these vectors, which will also be discussed.