International Mammalian Genome Society

The 13th International Mouse Genome Conference
October 31-November 3, 1999

Table of Contents * Structure * Bioinformatics * Sequence * Mapping * New Tools * Gene Discovery * Developmental * Mutagenesis * Functional Genomics

E56 The Use of Speed-Backcrosses for Updating the Harwell Mutant Map: A Novel Application for IVF Allows Rapid Generation of Large Numbers of Mice for Genotype Analysis

Christine Wells, Peter Glenister, Claire Thornton, Sarah Smythe and Steve Brown. Medical Research Council, Mammalian Genetics Unit, Harwell, OX11 0RD, UK

The success of the Harwell ENU mutagenesis programme has necessitated finding novel approaches to facilitate mapping the large numbers of interesting phenotypes already generated. For many years the major bottleneck for any mapping project has traditionally been the generation of sufficient numbers of backcross animals.

By the use of in vitro fertilisation we have obtained large numbers of animals in a very short space of time. Furthermore, this approach has cut demand on our animal facility by circumventing the traditional breeding route. More than 100 backcross progeny are routinely produced from one IVF session, the equivalent of 15 - 20 natural matings. The recipient foster females (~12 - 18) are housed in groups of 6 until just before parturition thus saving animal house space. A further advantage is that all the backcross animals are born and weaned on the same day. When the phenotype is classifiable, tail snips are frozen for genotype analysis and the animals removed from the shelf. IVF has proven so successful in this application that we are also able to archive lines by banking surplus embryos as well as freezing excess sperm for future IVF's if required.

Using this approach, 23 lines have been expanded for mapping in the initial 6 months. Using a multiplexed fluorescent genome scan we are aiming to map 20 lines within 10cM over the next year. Map positions have been obtained for 6 of these lines, representing several new loci. It now appears that the bottleneck has moved to the number of lines that can be screened; future work will be directed towards high throughput genomic analysis.


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