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

建立了逆转录病毒标记的小鼠胚胎干细胞库。在逆转录病毒载体中引入两组60粒条形码，通过对条形码序列的计数来估计细胞数量。首先，我们通过计算每个突变体的条形码读取数来检查该系统的准确性。这个读数几乎与我们事先得到的扩散速度一致。利用该系统，我们在体外分离了一些lincRNA突变体。我们分离了一个缺乏向中胚层分化的突变体。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。对克隆体的分析可能会找到如何防止植入的iPS细胞癌变的答案。

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.