School of Life Science and Technology, Wuhan Polytechnic University, Hubei, People's Republic of China.
State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Hubei, People's Republic of China.
CRISPR J. 2022 Feb;5(1):7-18. doi: 10.1089/crispr.2021.0039. Epub 2022 Jan 24.
The CRISPR-Cas nuclease has emerged as a powerful genome-editing tool in recent years. The CRISPR-Cas system induces double-strand breaks that can be repaired via the non-homologous end joining or homology-directed repair (HDR) pathway. Compared to non-homologous end joining, HDR can be used for the treatment of incurable monogenetic diseases. Therefore, remarkable efforts have been dedicated to enhancing the efficacy of HDR. In this review, we summarize the currently used strategies for enhancing the HDR efficiency of CRISPR-Cas systems based on three factors: (1) regulation of the key factors in the DNA repair pathways, (2) modulation of the components in the CRISPR machinery, and (3) alteration of the intracellular environment around double-strand breaks. Representative cases and potential solutions for further improving HDR efficiency are also discussed, facilitating the development of new CRISPR technologies to achieve highly precise genetic manipulation in the future.
近年来，CRISPR-Cas 核酸酶已成为一种强大的基因组编辑工具。CRISPR-Cas 系统诱导双链断裂，可通过非同源末端连接或同源定向修复 (HDR) 途径进行修复。与非同源末端连接相比，HDR 可用于治疗无法治愈的单基因疾病。因此，人们做出了巨大努力来提高 HDR 的效率。在这篇综述中，我们总结了目前基于三个因素提高 CRISPR-Cas 系统 HDR 效率的策略：(1) 调控 DNA 修复途径中的关键因素，(2) 调节 CRISPR 机器组件，以及 (3) 改变双链断裂周围的细胞内环境。还讨论了代表性案例和进一步提高 HDR 效率的潜在解决方案，为开发新的 CRISPR 技术以实现未来高度精确的基因操作奠定了基础。
