2013 Fiscal Year Final Research Report
Construdtion of Genome moving system utilizing a chromosome as transfer unit
| Project/Area Number |
22500426
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Tottori University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
INOUE Toshiaki 鳥取大学, 医学部, 准教授 (80305573)
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| Project Period (FY) |
2010-04-01 – 2013-03-31
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| Keywords | 微小核細胞 / 細胞融合 / 染色体導入 / 麻疹ウイルス / エンベロープタンパク質 / CD46 / 一本鎖抗体 / リターゲティング |
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| Research Abstract |
Microcell-mediated chromosome transfer allows moving a single chromosome from donor to recipient cells. A critical step determining the transfer efficiency is fusion of donor-derived microcells with recipient cells. We tested the fusogenic Hemmagulutinin (H) and Fusion (F) glycoproteins derived from an attenuated strain of Measles Virus (MV-Edm) for microcell fusion. Microcell hybrids were obtained with higher efficiency compared to the use of conventional fusogen Polyethylene Glycol. Yield of microcell hybrids was correlated with the level of cell surface expression in the recipient cells of CD46, which acts as receptor for MV. To achieve efficient fusion towards recipient cells such as fibroblasts with low level-expression of CD46, we tested the retargeting of microcells by adding scFv to the extracellular C-terminal end of the H protein. Addition of scFvs against CD13 and Transferrin receptor that are abundantly expressed in human fibroblasts improved chromosome transfer efficiency.
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