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。我们检测了麻疹病毒减毒株(MV-EDM)的融合血凝素(H)和融合(F)糖蛋白用于微细胞融合。与常规FusoGen聚乙二醇组相比，获得了效率更高的微细胞杂化材料。微细胞杂交种的产量与作为MV受体的CD46受体细胞表面表达水平相关。为了实现与CD46低表达的成纤维细胞等受体细胞的有效融合，我们通过在H蛋白的胞外C末端添加scFv来测试微细胞的重定向。加入在人成纤维细胞中大量表达的针对CD13和转铁蛋白受体的单链抗体可以提高染色体转移效率。

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.