Isolation and analysis of mutants that do not form teratoma using homo-mutagenized mouse ES cell library

使用同源诱变小鼠 ES 细胞库分离和分析不形成畸胎瘤的突变体

• 批准号: 24659180
• 负责人: YOSHIMURA Yasuhide
• 金额: $ 2.5万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24659180/
• 关键词: 再生医学; テラトーマ; 再生医療; lincRNA; 細胞分化

We established the homo-mutagenized mouse ES cell library labeled with retrovirus. Two sets of 60-mer barcode are introduced into the retrovirus vector and enable to estimate the cell number by counting of the barcode sequence. First we checked the accuracy of this system by counting the read number of barcode of each mutant. This read number is almost coincident with the proliferation speed that we got in advance. By using this system, we isolated some mutant of lincRNA in vitro. We isolated a mutant that had deficiency for differentiation to mesoblast. Using this clone as a marker, we will try to isolate the noble mutant that is not able to form teratoma. Analysis for the clone might lead to the answer how to prevent oncogenesis of implanted iPS cells.

我们建立了逆转录病毒标记的同种诱变小鼠ES细胞库。将两组60聚体条形码引入逆转录病毒载体，并通过对条形码序列的计数来估计细胞数量。首先，我们通过统计每个突变体的条形码读取次数来检查该系统的准确性。这个读数与我们事先得到的扩散速度几乎是一致的。利用该体系，我们在体外分离到了部分lincRNA突变体。我们分离到一株缺乏向中胚细胞分化的突变体。利用这个克隆作为标记，我们将尝试分离出不能形成畸胎瘤的高贵突变体。对克隆的分析可能会导致如何防止移植的iPS细胞发生肿瘤的答案。