International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


Greber B, Lehrach H, Himmelbauer H

Max Planck Institute for Molecular Genetics, Berlin, Germany

Despite the availability of mammalian genome sequences, the functions of many genes remain unknown. Mutant mice are valuable for studying mammalian gene functions and for modeling human disease phenotypes. With regard to the latter, a range of diseases involves the missplicing of pre-mRNAs. 

Embryonic stem cell technology offers the possibility to manipulate the mouse genome and select for mutations in vitro. Mutant animals can then be generated from selected clones. We are inducing mutations chemically using ENU, followed by the generation of a mutant clone library in a 96-well format. The library we have generated consists of about 40,000 clones distributed into 96 times 96 samples. Such clone libraries can be specifically screened for mutations in genes of interest. We have developed a protocol that allows the targeted detection of splice mutations based on highly pooled cDNA samples. The procedure does not require any high-cost instrumentation and is time-efficient enabling one worker to screen a given gene within about one week. As a proof of principle, we have isolated splice mutant clones for the mouse Kit gene and obtained germline transmission for a mutation causing the deletion of exon 18 at the transcript level. The heterozygous mutant displayed a dominant phenotype which resembled that of an established targeted null allele.

Acknowledgement of support: Our work is carried out through funding by the German National Genome Research Network (NGFN).

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