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

使用病毒载体一步式人类 iPS 细胞诱导和基因靶向

• 批准号: 22659061
• 负责人: MITANI Kohnosuke
• 金额: $ 1.98万
• 依托单位: Saitama Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22659061/
• 关键词: 相同組換え; iPS細胞; ウイルスベクター; 再生医療; 多能性幹細胞

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.

为了最小化用于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，我们研究了同时iPSC诱导和基因靶向的策略。通过使用仙台病毒载体SeVp（KOSM）对TIG 3人成纤维细胞进行iPSC诱导。iPSC样集落作为细胞群生长而不克隆，随后用靶向HPRT基因座的HDAdV转导。基因靶向频率为染色体整合的3-5%，这与真正的人iPSC的频率相当。从iPSC诱导开始到耐药菌落的分离，仅用了6周时间。

项目成果

One Step Human iPS Cell Induction And Enhanced Gene Targeting Using Viral Vectors And TALE Nucleases

使用病毒载体和 TALE 核酸酶一步完成人类 iPS 细胞诱导和增强基因靶向

• 发表时间: 2012
• 作者: Ko Mitani;Emi Aizawa;Claudio Mussolino;Manami Ohtaka;Ken Nishimura;Toni Cathomen;Mahito Nakanishi
• 通讯作者: Mahito Nakanishi

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

使用病毒载体一步式人类 iPS 细胞诱导和基因靶向

• 发表时间: 2012
• 作者: Ko Mitani;Emi Aizawa;Manami Ohtaka;Ken Nishimura;Mahito Nakanishi
• 通讯作者: Mahito Nakanishi