CRISPR/Cas介导的基因编辑在合成生物学中的应用：挑战与优化
The application of CRISPR /Cas mediated gene editing in synthetic biology: Challenges and optimizations.
作者信息
Li Wenqian, Huang Can, Chen Jingyu
机构信息
MOE Key Laboratory of Precision Nutrition and Food Quality, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.
Key Laboratory of Food Bioengineering (China National Light Industry), College of Food Science and Nutritional Engineering, China Agricultural University, Beijing , China.
出版信息
Front Bioeng Biotechnol. 2022 Aug 25;10:890155. doi: 10.3389/fbioe.2022.890155. eCollection 2022.
Clustered regularly interspaced short palindromic repeats (CRISPR) and its associated enzymes (Cas) is a simple and convenient genome editing tool that has been used in various cell factories and emerging synthetic biology in the recent past. However, several problems, including off-target effects, cytotoxicity, and low efficiency of multi-gene editing, are associated with the CRISPR/Cas system, which have limited its application in new species. In this review, we briefly describe the mechanisms of CRISPR/Cas engineering and propose strategies to optimize the system based on its defects, including, but not limited to, enhancing targeted specificity, reducing toxicity related to Cas protein, and improving multi-point editing efficiency. In addition, some examples of improvements in synthetic biology are also highlighted. Finally, future perspectives of system optimization are discussed, providing a reference for developing safe genome-editing tools for new species.
成簇规律间隔短回文重复序列（CRISPR）及其相关酶（Cas）是一种简单便捷的基因组编辑工具，在最近已被应用于各种细胞工厂和新兴的合成生物学中。然而，包括脱靶效应、细胞毒性和多基因编辑效率低在内的几个问题与CRISPR/Cas系统相关，这些问题限制了其在新物种中的应用。在本综述中，我们简要描述了CRISPR/Cas工程的机制，并基于其缺陷提出了优化该系统的策略，包括但不限于提高靶向特异性、降低与Cas蛋白相关的毒性以及提高多点编辑效率。此外，还强调了合成生物学中一些改进的例子。最后，讨论了系统优化的未来前景，为开发用于新物种的安全基因组编辑工具提供参考。
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