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

为了最大限度地减少用于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，我们研究了一种同时进行IPSC诱导和基因打靶的策略。用仙台病毒载体SEVP(KOSM)诱导TIG3人成纤维细胞IPSC。IPSC样菌落作为细胞群体生长，不进行克隆，随后用针对HPRT基因的HDAdV进行转导。基因打靶频率为染色体整合率的3-5%，与真人IPSCs的打靶频率相当。从IPSC诱导到耐药克隆的分离，仅用了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.