Gene Editing Tools Based on CRISPR/Cas System and Their Improvement Strategies

doi:10.13523/j.cb.2303031

China Biotechnology

2023, Vol. 43

Issue (8): 72-85 DOI: 10.13523/j.cb.2303031

Gene Editing Tools Based on CRISPR/Cas System and Their Improvement Strategies

XIN Yuan¹,TIAN Kai-ren³,QIAO Jian-jun^1,^2,³,CAIYIN Qing-ge-le^1,^2,^**(

)

1 School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
2 Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China
3 Zhejiang Institute of Tianjin University (Shaoxing), Shaoxing 312300, China

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Abstract

CRISPR/Cas system has continuously promoted the progress in the field of life science since its discovery. At the same time, the gene editing technology mediated by CRISPR/Cas is also developing and improving continuously. The development of new gene editing tools such as CRISPR/Cas gene editing technology based on DSBs repair, base editors and prime editors has paved the way for basic biological research. Although these tools have brought revolutionary changes to biotechnology, problems such as low efficiency of gene editing, low purity of products and frequent off-target effects began to emerge. The continuous development of accurate, efficient, and secure CRISPR/Cas editing tools remains a current and future research focus. Here, the development, composition and principle of CRISPR/Cas system based gene editing tools are briefly described. The general strategies for the CRISPR/Cas system to improve editing efficiency, expand editing scope and reduce off-target effects are systematically summarized, as well as the improvement methods of different CRISPR/Cas gene editing tools. In addition, the future research direction of CRISPR gene editing tools is discussed.

Key words： CRISPR/Cas Gene editing efficiency DSBs Base editors Prime editors

Received: 10 March 2023 Published: 05 September 2023

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Cite this article:

XIN Yuan, TIAN Kai-ren, QIAO Jian-jun, CAIYIN Qing-ge-le. Gene Editing Tools Based on CRISPR/Cas System and Their Improvement Strategies. China Biotechnology, 2023, 43(8): 72-85.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2303031 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I8/72

Fig.1 Principle of CRISPR/Cas gene editing system (a) The gene editing principle of Cas9 and Cas12 (b) The gene editing principle of base editings (c) The gene editing principle of prime editings

Fig.2 gRNA engineering (a) Optimization of structure (b) RNA array for multi-gene editing (c) Chemical modification (d) Introducing functional structures

Table 1 High-fidelity variants of Cas proteins

Table 2 Cas9 orthologs and their variants with expanded targeting scope

Table 3 Cas12 orthologs and their variants with expanded targeting scope

Fig.3 Optimization strategies for base editors and prime editors


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