International Mammalian Genome Society

17th International Mouse Genome Conference

9-12 November 2003, Braunschweig, Germany


POSTER 61 - ESTABLISHMENT OF A SCREENING SYSTEM FOR MUTANTS AFFECTING GENOMIC IMPRINTING IN THE RIKEN-GSC PROJECT

Suzuki T
Mouse Functional Genomics Research Group, Genomics Science Center (GSC), RIKEN Yokohama Institute

Co-Authors: 2) Furuumi H, 3) Hashimoto M, 1) Kaneda H, 4) Gondo Y, 1) Noda T, 1) Wakana S, 5) Ishino F, 2) Sasaki H, 1) Shiroishi T
Institutions: 1) Mouse Functional Genomics Research Group, Genomics Science Center (GSC), RIKEN Yokohama Institute, 2) Department of Integrated Genetics, National Institute of Genetics, 3) Gene Research Center, Tokyo Institute of Technology, 4) Population and Quantitative Genomics Team, GSC, RIKEN, 5) Department of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University

Genomic imprinting is epigenetic regulation of gene expression depending on parental origin of the genes in mammals. Proper regulation of the imprinted genes is essential for normal development, and a number of human disorders are associated with disturbance of the genomic imprinting. Initiation process of the genomic imprinting during gametogenesis consists of two stages, erasure of the parental imprints and re-establishment of sex-different imprints. These two stages frequently coincide with change of methylation status of the imprinted genes, but very little is known about the initiation mechanism of the genomic imprinting.

In order to detect and identify novel genes involved in the initiation of the genomic imprinting, we established screening system for mutants affecting the genomic imprinting with ENU-mutagenised mice in the RIKEN GSC program. In the screening, we measure methylation level of the imprinted genes in the mature gametes using methylation sensitive enzymes or the COBRA (combined bisulfite restriction analysis). To detect the mutants affecting the two stages, we assay the methylation status of the two kinds of imprinted genes. The paternally methylated gene H19 is subjected to the assay in screening for the re-establishment stage during spermatogenesis and for the erasure stage during oogenesis. In reverse, the maternally methylated genes, Igr2r, Peg1, and Peg3, are subjected to the assay in screening for the re-establishment stage during oogenesis and for the erasure stage during spermatogenesis. We have started screening for mutants affecting genomic imprinting using this protocol, and report the current results of the screening.


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