Construdtion of Genome moving system utilizing a chromosome as transfer unit

微细胞介导的染色体转移允许将单个染色体从供体细胞转移到受体细胞。决定转移效率的关键步骤是供体来源的微细胞与受体细胞的融合。我们测试了源自麻疹病毒减毒株（MV-Edm）的促融合血谷凝集素（H）和融合（F）糖蛋白用于微细胞融合。与使用常规的融合剂聚乙二醇相比，以更高的效率获得微细胞杂交体。微细胞杂交体的产量与受体细胞中作为MV受体的CD 46的细胞表面表达水平相关。为了实现与受体细胞如具有低水平表达的CD 46的成纤维细胞的有效融合，我们通过将scFv添加到H蛋白的细胞外C末端来测试微细胞的重靶向。添加针对在人成纤维细胞中大量表达的CD 13和转铁蛋白受体的scFv改善了染色体转移效率。

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.