人类细胞中的基因组工程。

Genome engineering in human cells.

作者信息

Song Minjung, Kim Young-Hoon, Kim Jin-Soo, Kim Hyongbum

机构信息

Graduate School of Biomedical Science and Engineering, College of Medicine, Hanyang University, Seoul, South Korea.

Center for Genome Engineering, Institute for Basic Science, Seoul, South Korea; Department of Chemistry, Seoul National University, Seoul, South Korea.

出版信息

Methods Enzymol. 2014;546:93-118. doi: 10.1016/B978-0-12-801185-0.00005-2.

DOI:10.1016/B978-0-12-801185-0.00005-2

PMID:25398337

Abstract

Genome editing in human cells is of great value in research, medicine, and biotechnology. Programmable nucleases including zinc-finger nucleases, transcription activator-like effector nucleases, and RNA-guided engineered nucleases recognize a specific target sequence and make a double-strand break at that site, which can result in gene disruption, gene insertion, gene correction, or chromosomal rearrangements. The target sequence complexities of these programmable nucleases are higher than 3.2 mega base pairs, the size of the haploid human genome. Here, we briefly introduce the structure of the human genome and the characteristics of each programmable nuclease, and review their applications in human cells including pluripotent stem cells. In addition, we discuss various delivery methods for nucleases, programmable nickases, and enrichment of gene-edited human cells, all of which facilitate efficient and precise genome editing in human cells.

摘要

人类细胞中的基因组编辑在研究、医学和生物技术领域具有重要价值。包括锌指核酸酶、转录激活样效应因子核酸酶和RNA引导的工程核酸酶在内的可编程核酸酶能够识别特定的靶序列，并在该位点产生双链断裂，这可能导致基因破坏、基因插入、基因校正或染色体重排。这些可编程核酸酶的靶序列复杂度高于单倍体人类基因组的大小——320万个碱基对。在此，我们简要介绍人类基因组的结构以及每种可编程核酸酶的特性，并综述它们在包括多能干细胞在内的人类细胞中的应用。此外，我们还讨论了核酸酶、可编程切口酶的各种递送方法以及基因编辑人类细胞的富集，所有这些都有助于在人类细胞中进行高效且精确的基因组编辑。