利用 Gibson 组装法制备供体载体对人细胞系进行 CRISPR-Cas9 基因组编辑。
CRISPR-Cas9 Genome Editing in Human Cell Lines with Donor Vector Made by Gibson Assembly.
机构信息
Department of Biology, University of Texas - Rio Grande Valley, Edinburg, TX, USA.
出版信息
Methods Mol Biol. 2020;2115:365-383. doi: 10.1007/978-1-0716-0290-4_20.
CRISPR Cas9 genome editing allows researchers to modify genes in a multitude of ways including to obtain deletions, epitope-tagged loci, and knock-in mutations. Within 6 years of its initial application, CRISPR-Cas9 genome editing has been widely employed, but disadvantages to this method, such as low modification efficiencies and off-target effects, need careful consideration. Obtaining custom donor vectors can also be expensive and time-consuming. This chapter details strategies to overcome barriers to CRISPR-Cas9 genome editing as well as recent developments in employing this technique.
CRISPR Cas9 基因组编辑允许研究人员以多种方式修改基因，包括获得缺失、表位标记基因座和基因敲入突变。在最初应用的 6 年内，CRISPR-Cas9 基因组编辑已经得到了广泛的应用，但该方法存在一些缺点，例如修饰效率低和脱靶效应，需要仔细考虑。获得定制的供体载体也可能昂贵且耗时。本章详细介绍了克服 CRISPR-Cas9 基因组编辑障碍的策略，以及利用该技术的最新进展。
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