International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


POSTER 87 - EXPRESSION GENE TRAP MUTAGENESIS AT THE CENTRE FOR MODELING HUMAN DISEASE (CMHD)

A Johnson
University of Toronto

1) Lan Q, 1) Li CYJ, 1) Ohishi M, 1) Reid T, 2) To C, 2)Tsao N, 1) Yu M, 2) Kassam N, 1) Osborne L, 2) Rossant J, 1) Stanford WL, and the CMHD
1) University of Toronto; 2) Samuel Lunenfeld Research Institute

We are performing gene trap-based expression and genotypic screens to generate new mouse mutations that will help delineate the molecular controls of specific developmental programs. Random insertion of the gene trap vectors containing a splice acceptor immediately upstream of a promoterless reporter into the mouse embryonic stem (ES) cell genome provides a method to generate loss-of-function mutations and report expression for many, and given new vectors potentially all, mouse genes. A series of polyA trap vectors have been developed to attempt to optimize insertions into all classes of genes, irrespective of their expression in undifferentiated ES cells. The gene trap insertions are screened using multiplexed in vitro differentiation and induction assays. An expression profile for each clone is being generated being generated and will be available via an online searchable database by the time of the International Mammalian Genome Conference (http://www.cmhd.ca/sub/genetrap.htm). RACE-PCR is being performed to complement expression profiles with sequence tags, which will eventually be available online as well. The sequence tags will also be submitted to the NCBI gene trap database where they will be linked to their physical map positions and The International Gene Trap Consortium database being developed. The clones will be available as a resource to researchers worldwide to help functionally annotate the mammalian genome and will serve as a source to test candidate loci identified by phenotype-driven mutagenesis screens.


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