| Project/Area Number |
17016014
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SAITO Izumu The University of Tokyo, 医科学研究所, 教授 (70158913)
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| Co-Investigator(Kenkyū-buntansha) |
KANEGAE Yumi 東京大学, 医科学研究所, 助教 (80251453)
KONDO Saki 東京大学, 医科学研究所, 助教 (80451871)
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| Project Period (FY) |
2005 – 2009
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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥128,600,000 (Direct Cost: ¥128,600,000)
Fiscal Year 2009: ¥25,500,000 (Direct Cost: ¥25,500,000)
Fiscal Year 2008: ¥25,500,000 (Direct Cost: ¥25,500,000)
Fiscal Year 2007: ¥25,500,000 (Direct Cost: ¥25,500,000)
Fiscal Year 2006: ¥25,500,000 (Direct Cost: ¥25,500,000)
Fiscal Year 2005: ¥26,600,000 (Direct Cost: ¥26,600,000)
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| Keywords | アデノウイルスベクター / 遺伝子治療 / 部位特異的組換え酵素 / Cre / loxP / FLP / FRT / guttedベクター / アデノウイスルベクター |
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| Research Abstract |
Aiming gene therapy for "unvisible cancer" such as disseminating or transferring malignant microtumors, we developed a "single-type, cancer-specific adenovirus vector with high-level expression" possessing cell specificity and high expression efficiency. For addition of high specificity, we used a cancer-specific promoter. Construction methods of two vector systems, a "stuffer-deletion type" and a novel "excisional-expression type", were established. Each system contains two expression units: a "switch unit" expressing Cre under the control of a cell-specific promoter with very low activity, and a "target unit" expressing under the control of very high activity depending on the Cre activity. During the process, it was found that Cre was expressed at a leak and influenced on production of the purpose vector. Therefore, we produced and screened dominant negatives and shRNA against Cre. Using them, we succeeded in the development of high-purity production of the "excisional-expression type" vector. Because the vector established here showed high expression efficiency selectively to cancer cells, we then established a model muse system for examining effective treatment to disseminated cancers. In addition, we also analyzed RMCE (recombinase-mediated cassette exchange) reaction of Cre and FLP, and succeeded in establishment of novel production method, which enabled us to produce many vectors simultaneously.
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