International Mammalian Genome Society

The 15th International Mouse Genome Conference (2001)


POSTER 227 - THE IDENTIFICATION OF ENU INDUCED MUTANTS IN THE MOUSE WITH RELEVANCE TO TYPE II DIABETES USING A GENE DRIVEN APPROACH

Alison Hugill

Medical Research Council

Oxford

OX11 0RD

UK

1)Coghill E., 2)Hunter A.J., 1)Brown S.D.M., 1)Cox R.D. 

1)MRC Mammalian Genetics Unit and Mouse Genome Centre

2)GlaxoSmithKline, New Frontiers Science Park

 

N-ethyl-N-nitrosourea mutagenesis (ENUm) programmes in mice provide a route to gene driven approaches where mutations in known genes can be recovered. We have generated a parallel DNA/sperm archive of ENUm mice that can be rapidly screened for mutations in the DNA followed by recovery of mutants from the sperm bank. ENUm is complementary to targeted mutation technologies in that a large variety of protein alterations with different phenotypes can be generated. We have established 2,230 DNA/sperm samples that with a specific locus mutation rate of 0.0015 would give a >90% chance of recovering 1 allele at any locus and an 80% chance of recovering 2 alleles. As part of an EU project we are constructing a new archive of DNA/sperm from BALB/c x C3H F1 ENUm mice. The project aim is to generate an archive of approx. 5,000 DNAs that may allow an allelic series of 3 or more alleles to be obtained with >90% confidence. We are currently carrying out a pilot screen of the existing archive with 3 candidate genes that are important in glucose homeostasis and could play a role in Type II Diabetes. These are p85a, the catalytic subunit of phosphoinositide 3-kinase, Resistin, and Peroxisome proliferator activated receptor gamma. The results of this screen will be presented. The study is being carried out using Denaturing High Performance Liquid Chromatography on a Transgenomic Wave machine to scan for heterozygous mutations.


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