The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
Nat Biotechnol. 2011 Jul 7;29(8):731-4. doi: 10.1038/nbt.1927.
Targeted genetic engineering of human pluripotent cells is a prerequisite for exploiting their full potential. Such genetic manipulations can be achieved using site-specific nucleases. Here we engineered transcription activator-like effector nucleases (TALENs) for five distinct genomic loci. At all loci tested we obtained human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) clones carrying transgenic cassettes solely at the TALEN-specified location. Our data suggest that TALENs employing the specific architectures described here mediate site-specific genome modification in human pluripotent cells with similar efficiency and precision as do zinc-finger nucleases (ZFNs).
靶向基因工程人类多能细胞是开发其全部潜力的前提。这种基因操作可以使用位点特异性核酸酶来实现。在这里,我们设计了针对五个不同基因组位点的转录激活因子样效应物核酸酶(TALENs)。在所有测试的位点上,我们获得了携带转基因盒的人胚胎干细胞(ESC)和诱导多能干细胞(iPSC)克隆,这些克隆仅在 TALEN 指定的位置上。我们的数据表明,采用这里描述的特定结构的 TALEN 介导人多能细胞的特异性基因组修饰,其效率和精度与锌指核酸酶(ZFNs)相似。