From the Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037 and.
J Biol Chem. 2014 Feb 21;289(8):4594-9. doi: 10.1074/jbc.R113.488247. Epub 2013 Dec 20.
Human pluripotent stem cells (hPSCs) offer unprecedented opportunities to study cellular differentiation and model human diseases. The ability to precisely modify any genomic sequence holds the key to realizing the full potential of hPSCs. Thanks to the rapid development of novel genome editing technologies driven by the enormous interest in the hPSC field, genome editing in hPSCs has evolved from being a daunting task a few years ago to a routine procedure in most laboratories. Here, we provide an overview of the mainstream genome editing tools, including zinc finger nucleases, transcription activator-like effector nucleases, clustered regularly interspaced short palindromic repeat/CAS9 RNA-guided nucleases, and helper-dependent adenoviral vectors. We discuss the features and limitations of these technologies, as well as how these factors influence the utility of these tools in basic research and therapies.
人类多能干细胞(hPSCs)为研究细胞分化和人类疾病模型提供了前所未有的机会。精确修饰任何基因组序列的能力是实现 hPSCs 全部潜力的关键。由于 hPSC 领域的巨大兴趣推动了新型基因组编辑技术的快速发展,hPSC 中的基因组编辑已经从几年前的一项艰巨任务发展成为大多数实验室的常规程序。在这里,我们概述了主流的基因组编辑工具,包括锌指核酸酶、转录激活因子样效应物核酸酶、成簇规律间隔短回文重复/ Cas9 RNA 引导的核酸酶和辅助依赖性腺病毒载体。我们讨论了这些技术的特点和局限性,以及这些因素如何影响这些工具在基础研究和治疗中的应用。