International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)

H20. A Noble Strategy for Mutagenesis in the Mouse: Heavy Ion Beams and Fetal Phenotype Screening -----I. An Overview

Moriaki Kusakabe1, Atsushi Yoshiki1, Noriko Hiraiwa1, Fumio Ike1, Chikako Yoshida-Noro1, Hiroki Nagase2 and Yasushige Yano3
1Experimental Animal Research Division, and
Molecular Oncology, RIKEN Tsukuba Institute, Tsukuba, Ibaraki 305-0074, Japan;
Cyclotron Laboratory, RIKEN, Wako, Saitama 351-0198, Japan

Mouse mutant archive is a useful resource for the functional genomics in the post-genomic era. We propose here noble systematic approaches for the production, screening, preservation and analysis of mouse mutants.

1.Heavy Ion Beam Mutagenesis

Since on-going ENU chemical mutagenesis primarily induces random, single base pair changes, mutations in the genome do not necessarily represent as phenotypes. In order to increase the efficiency of phenotype assessment, we introduced a method using heavy ion beam generated from RIKEN ring cyclotron facilities as a mutagen. As this method is expected to cause deletions and mutations of the gene cluster, it is suitable for the analysis of multi-gene phenotypes such as cancer or developmental defects.

2. Fetal Phenotype Screening

Fertilization rate and birth rate are the key factors for getting mutant offspring, which causes difficulty in an effective accumulation of the mutation resource through the analysis of the born offspring. In order to overcome this problem, we apply the phenotype screening at the fetal stage for the developmental abnormalities.

3. Analysis for Expression Profiles in the Mutant Tissues

Gene expression profiles of the abnormal tissues in the mutant are analyzed by using Micro-Tissue cDNA Subtraction Library and in situ hybridization with a high-throughput system. By combining these data with genome analysis such as SSLP, RLGS or RDA, we study the mechanisms of which the mutated gene and resulted changes of gene cascade generate the mutant phenotype.

4. Preservation and Functional Analysis using EG cells

Primordial germ cells are isolated from the genital ridges in the fetus with mutant phenotype and cultured for establishing embryonic germ (EG) cells. Mutant genetic resource is cryopreserved in the form of EG cells and utilized by the production of chimeric or clone mice. Function of lethal gene mutation in the live animal can be examined by chimeric analysis.

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