1997 Fiscal Year Final Research Report Summary
A gene trap approach to identify a new gene involved in central nervous system development
| Project/Area Number |
07557092
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | Niigata University |
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Principal Investigator |
KUWANO Ryozo Niigata University, Research Laboratory for Molecular genetics, Associate Professor, 遺伝子実験施設, 助教授 (20111734)
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| Co-Investigator(Kenkyū-buntansha) |
KUMANISHI Toshiro Niigata University, Brain Research Institute, Professor, 脳研究所, 教授 (40018601)
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| Project Period (FY) |
1995 – 1997
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| Keywords | gene / cell abration / Cre-lox / diphteria toxin / gene trapping / toransgenic mice / brain / immunohistochemistry |
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| Research Abstract |
In attempt to elucidate of interaction between neuron and glia involved in neuronal function we used a strategy of cell ablation using a diphtheria toxin A fragment (DT-A) gene in transgenic mice. Based on spatial-temporal specific gene expression using an appropriate promoter such as S100 or GFAP promoter in astrocyte and a novel one for neuron discovered by gene trapping, the DT-A gene is activated in the restricted cells in the central nervous system. In addition we introduced cre-lox system for regional and stage specific regulation of the target gene. expression. Cre-recombinase mediated site specific recombination occurs on the restricted DNA sequence composed of 34 bp(loxP). DT-A activity was suppressed by insertion of transcription termination signal down stream of murine molony leukemia virus promoter. Cell ablation by cre-lox has to be verified in culture system prior to create transgenic mice. We confirmed an effect of DT-A in the transfected cells. We produced several transgenic mouse lines of which some showed ectopic expression pattern owing to a position effect.
Gene trapping provided a novel promoter expressed in the central nervous system and heart. A new construct of the target gene containing insulator sequence for cell ablation is constructed again.
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