基于CRISPR/Cas9技术的基因敲入/敲除策略

成簇的规律间隔性短回文序列(CRISPR)基因编辑系统,因其设计简单操作方便和无种属限制,已成为一种广泛应用的基因组定点编辑工具,在复杂的基因组编辑,例如基因的人源化改造以及条件等位基因的构建中有所应用。在自然界中,CRISPR系统拥有多种类别。其中,CRISPR/Cas9系统是研究最深入、应用最成熟的一种。本文针对CRISPR/Cas9系统,分别从基因敲入/敲除片段的大小、同源臂长短、构型即递送方式等技术环节进行综述,阐述不同设计及操作条件下由CRISPR/Cas9系统介导的基因敲入/敲除的效率差异。

Strategy of Gene Knockin/Knockout Based on CRISPR/Cas9 Technology

CRISPR (cluster regularly interspaced short palindrome repeats) gene editing system has become a widely used genome editing tool because of its remarkable advantages such as relatively easy in design and production, as well as no species limit. It has been used in complex genome editing, such as humanization of genes and the construction of conditional alleles. CRISPR system has a variety of categories in nature. CRISPR/Cas9 system is one of the most in-depth studies and the most mature technologies. In this review, we mainly focus on the important structure features of CRISPR/Cas9, such as gene editing fragment, length of homologous arms and its corresponding structure, as well as its delivery methods. The difference in efficiency of gene knockin/knockout mediated by the CRISPR/Cas9 system under different design and operating conditions was also summarized.