Hendriks William T, Warren Curtis R, Cowan Chad A
The Collaborative Center for X-Linked Dystonia Parkinsonism, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Brain Science Initiative, Harvard Medical School, Boston, MA 02114, USA.
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA.
Cell Stem Cell. 2016 Jan 7;18(1):53-65. doi: 10.1016/j.stem.2015.12.002.
Human pluripotent stem cells (hPSCs) with knockout or mutant alleles can be generated using custom-engineered nucleases. Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 nucleases are the most commonly employed technologies for editing hPSC genomes. In this Protocol Review, we provide a brief overview of custom-engineered nucleases in the context of gene editing in hPSCs with a focus on the application of TALENs and CRISPR/Cas9. We will highlight the advantages and disadvantages of each method and discuss theoretical and technical considerations for experimental design.
利用定制工程核酸酶可以生成带有敲除或突变等位基因的人类多能干细胞（hPSC）。转录激活样效应核酸酶（TALEN）和规律成簇间隔短回文重复序列（CRISPR）-Cas9核酸酶是编辑hPSC基因组最常用的技术。在本方案综述中，我们简要概述了在hPSC基因编辑背景下的定制工程核酸酶，重点是TALEN和CRISPR/Cas9的应用。我们将强调每种方法的优缺点，并讨论实验设计的理论和技术考量。
