| Project/Area Number |
12558087
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Nerve anatomy/Neuropathology
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| Research Institution | OSAKA CITY UNIVERSITY (2001)
Asahikawa Medical College (2000) |
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Principal Investigator |
KIYAMA Hiroshi Osaka City University, Graduate School of medicine, Professor, 大学院・医学研究科, 教授 (00192021)
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| Co-Investigator(Kenkyū-buntansha) |
NAMIKAWA Kazuhiko Osaka City University, Graduate School of medicine, Research Associate, 大学院・医学研究科, 助手 (50312468)
SEO Sumiko Osaka City University, Graduate School of medicine, Research Associate, 大学院・医学研究科, 助手 (70311529)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 2001: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2000: ¥8,200,000 (Direct Cost: ¥8,200,000)
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| Keywords | adenovrus / Cre-loxP / Promoter / NRSE / neuron specific / Gene transfer |
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| Research Abstract |
We have attempted to establish the adenovirus-mediated gene-transfer system, which enables neural species-specific expression. For this aim, Cre-loxP system was employed. Neuron, astrocyte, or Schwann cell specific promoters were examined in vitro. For the neuron specific expression, the basic promoter for SCG10 was used, and additional neuron specific silencer elements (NRSE) were attached. This modification increased the neuron specificity significantly. For the astrocyte expression, GFAP promoter was selected. Those promoters were used for the expression of Cre, and additional adenovirus vector, which encodes loxP-Stuffer-polyA-loxP-EGFP sequence downstream of a strong general promoter, was constructed. By the co-infection of these two adenoviruses, neural specie specific expression was attempted. The promoters we constructed for neurons and astrocytes worked well, and showed very high selectivity in culture such as the brain derived primary culture. Next we examined the specificity of the expression in rat brain. The combination of the adenovirus vectors was injected into various brain regions. The selectivity and expression level of the neuron specific promoter was nice, however the minor differences in the expression level among the injected regions were observed. This difference may be due to the core promoter of SCG10 ; nevertheless the selectivity in various regions was good. For the astrocyte specific expression, both selectivity and expression level was relatively good, but a few neuronal cells expressing EGFP was also found. However the expression level in neuronal cells were low. We have not obtained the Schwann cell specific promoter yet, and the study is on going.
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