| Project/Area Number |
63490020
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | School of Medicine, Tokai University |
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Principal Investigator |
KIMURA Minoru Tokai Univ. School of Med. Associate Prof., 医学部, 助教授 (10146706)
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| Co-Investigator(Kenkyū-buntansha) |
UEYAMA Yoshito Tokai Univ. School of Med. Associate Prof., 医学部, 助教授 (30072408)
YOKOYAMA Minesuke Central Institute for Experimental Animals Researcher, 研究員 (40090930)
KATSUKI Motoya Tokai Univ. School of Med. Prof., 医学部, 教授 (20051732)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1988: ¥4,300,000 (Direct Cost: ¥4,300,000)
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| Keywords | Transgenic mice / Antisense DNA / Genetic engineering / Embryo manipulation / Model for diseases / Myelin formation / 遺伝子発現 / ミエリン塩基性タンパク質 / 疾患モデル動物 |
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| Research Abstract |
The aim of this project is to construct the transgenic mice which reduce the expression of the target gene using antisense DNA and to reveal the exact function of the individual gene in vivo based on the phenotype of these transgenic mice.
Recently, we have developed this system for the myelin basic protein (MBP) gene. In the transgenic mice which expressed antisense MBP RNA (minus-strand RNA complimentary to a MBP mRNA), the endogeneous MBP mRNA, the MBP and the myelination in the central nervous system were reduced and consequently shiverer phenotype appeared (Science 241, 593-595 (1988)). Further analysis revealed that the reduction of MBP mRNA and MBP occurred around two weeks after birth when MBP expression and myelin formation of normal mice become active. The reduction level was correlated with the extent of shivering phenotype. Moreover, all kinds of the antigenically-related MBPs were simultaneously reduced in the antisense MBP transgenics, although the antisense cDNA used corresponded to the single kind of several MBP mRNA produced by alternative splicing.
To apply this system for other genes, we have made the antisense cDNA constructs of three genes (beta-actin, c-Ha-ras and dystrophin genes) under the appropriate promoters and produced the transgenic mice. Total 31 transgenic lines were established but no prominent phenotype were recognized until now. The expression of the antisense transgene was very low in the several transgenic mice so far examined.
Together with extensive analyses of these transgenic mice, we are now improving the structure of the introduced DNA to increase the antisense RNA expression.
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