International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


Y Sakuraba
Population and Quantitative Genomics Team, RIKEN Genomic Sciences Center

1) Sezutsu H, 1) Takahasi KR, 2) Inoue M, 2) Kaneda H, 2) Minowa O, 2) Wakana S, 2) Noda T, 2) Shiroishi T, 1) Gondo Y
1) Population and Quantitative Genomics Team and 2) Mouse Functional Genomics Research Group, RIKEN Genomic Sciences Center

Many inheritable mutants have been isolated by dominant screening in RIKEN ENU mutagenesis ( In the mutagenesis project, it is crucial to identify the site of mutations in the genome for the mutants. To this purpose we are currently taking three major approaches: Positional cloning. ENU-mutagenized C57BL/6 males are mated to DBA/2 females to produce G1 for the phenotype-driven approach. Phenodeviant G1 are subjected to inheritance test in G2 progeny that also provide linkage for the rough mapping. The sites of mutations will be identified by fine mapping and positional cloning or candidate gene approach as described below. Candidate gene approach. It is now plausible to list up candidate genes around the rough-mapped region due to the substantial public database of mouse and/or human genome. We are now designing PCR primers for direct-sequencing of candidate genes of some mutants. We have found three possible causable point mutations in Pax-6 gene. The primers that are prepared for the candidate gene approach are also useful for the following approach. Gene-driven approach. All the sperms of G1 males have been cryopreserved in liquid nitrogen to retain any mutations. The genomic DNAs from all the G1 males are extracted from the testes. Using these genetic resources, we have been studying the feasibility of the gene-driven mutagenesis. To detect the site of mutations in target genes with high-resolution and high-throughput manner, we have been trying to develop some primary screening methods by using TGCE and DHPLC in addition to the direct sequencing method.

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