School of Chemical, Materials and Biomedical Engineering, College of Engineering, University of Georgia, Athens, GA 30602, USA.
School of Chemical, Materials and Biomedical Engineering, College of Engineering, University of Georgia, Athens, GA 30602, USA.
Trends Biotechnol. 2024 Jan;42(1):104-118. doi: 10.1016/j.tibtech.2023.06.012. Epub 2023 Jul 26.
Microbial cell factories (MCFs) convert low-cost carbon sources into valuable compounds. The CRISPR/Cas9 system has revolutionized MCF construction as a remarkable genome editing tool with unprecedented programmability. Recently, the CRISPR toolbox has been significantly expanded through the exploration of new CRISPR systems, the engineering of Cas effectors, and the incorporation of other effectors, enabling multi-level regulation and gene editing free of double-strand breaks. This expanded CRISPR toolbox powerfully promotes MCF construction by facilitating pathway construction, enzyme engineering, flux redistribution, and metabolic burden control. In this article, we summarize different CRISPR tool designs and their applications in MCF construction for gene editing, transcriptional regulation, and enzyme modulation. Finally, we also discuss future perspectives for the development and application of the CRISPR toolbox.
微生物细胞工厂（MCFs）将低成本碳源转化为有价值的化合物。CRISPR/Cas9 系统作为一种具有空前可编程性的卓越基因组编辑工具，彻底改变了 MCF 的构建。最近，通过探索新的 CRISPR 系统、Cas 效应因子的工程设计以及其他效应因子的引入，CRISPR 工具包得到了显著扩展，实现了无需双链断裂的多层次调控和基因编辑。这个扩展的 CRISPR 工具包通过促进途径构建、酶工程、通量再分配和代谢负担控制，有力地推动了 MCF 的构建。在本文中，我们总结了不同的 CRISPR 工具设计及其在 MCF 构建中的应用，用于基因编辑、转录调控和酶调节。最后，我们还讨论了 CRISPR 工具包的未来发展和应用前景。
