ONE STEP HUMAN iPS CELL INDUCTION AND GENE TARGETING USING VIRAL VECTORS

为了减少iPSC诱导和基因修复的细胞培养，从而降低引入新突变的风险，我们研究了同时诱导iPSC和基因靶向的策略。使用仙台病毒载体SeVp（KOSM）诱导TIG3人成纤维细胞进行iPSC诱导。ipsc样菌落在没有克隆的情况下作为细胞群生长，随后用靶向HPRT位点的HDAdV进行转导。基因靶向频率为染色体整合的3-5%，与真正的人iPSCs相当。从诱导多能干细胞开始到耐药菌落的分离，只花了6周时间。

In order to minimize cell culture for iPSC induction and gene repair, thus reducing the risk of introducing new mutations, we examined a strategy of simultaneous iPSC induction and gene targeting. TIG3 human fibroblasts were subjected to iPSC induction by using the Sendai virus vector, SeVp(KOSM). iPSC-like colonies were grown as a cell populationwithout cloning, and were subsequently transduced with a HDAdV targeting the HPRT locus. The gene targeting frequencies were 3-5% of chromosomal integration, which were equivalent to those with authentic human iPSCs. From the start of iPSC induction to isolation of drug-resistant colonies, it took only 6 weeks.