International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)

F6. Development of an Oligonucleotide Array for Monitoring Upregulated Genes in a Mouse GVHR Model

Masatoshi Wakui1, Katsushi Tokunaga1, Yoko Hatta2, Nobuhiko Morimoto2, Sachiko Karaki2, Tohru Makino2, Ken-ichi Kurata3, Akira Suyama3
1Department of Human Genetics, Graduate School of Medicine, University of Tokyo,
Life-science Technology Research Center, Olympus Optical Co.,LTD,
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Tokyo, Japan

Graft-versus-host reaction (GVHR), which is a major immunological complication after allogeneic hematopoietic stem cell transplantation and transfusion, is a complex molecular process initiated upon allorecognition. To identify genes relevant to the pathogenesis of GVHR, we performed the differential display method (DD) for in vivo specimens in murine transplant models, and identified upregulated genes in the early phase of murine GVHR. They included six known genes, such as an interferon (IFN)-gamma-induced 47 kDa GTPase, a mammalian septin, or a putative mouse homologue of a human cancer-related gene, and one uncharacterized gene corresponding to a mouse expressed sequence tag (EST). These genes and the related genes could present candidates for molecular targets of clinical application as well as a framework for functional studies on the molecular pathogenesis of GVHR. For further investigation of gene expression profiles in murine GVHR, we have been developing a novel oligonucleotide array system. A total of 33 genes, whose sequences are available in the public databases, were selected as potential targets for monitoring murine GVHR at the trnanscription level. They belong to several gene families (IFN-gamma-induced 47 kDa and 65-k Da GTPases, mammalian septins, and activin members) or functionally related gene groups (extracellular matrix components and oxidative stress-related molecules). On the array, 30 mer oligonucleotides are used as specific probes for detection of each gene expression. The design for the probes was carried out using a newly developed efficient algorithm for finding out a set of unique sequences of target transcripts with uniform melting temperature and no folding potential. Towards the clinical application, the assessment of the efficacy of this novel array system on gene expression profiling is ongoing. This work was perfomed as a part of the research and development project of Industrial Science and Technology Program supported by NEDO (New Energy and Industrial Technology Development Organization).


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