A highly effective metabolic pathway is the key for an efficient cell factory. However, the engineered homologous or heterologous multi-gene pathway may be unbalanced, inefficient and causing the accumulation of potentially toxic intermediates. Therefore, pathways must be constructed optimally to minimize these negative effects and maximize catalytic efficiency. With the development of CRISPR technology, some of the problems of previous pathway engineering and genome editing techniques were resolved, providing higher efficiency, lower cost, and easily customizable targets. Moreover, CRISPR was demonstrated as robust and effective in various organisms including both prokaryotes and eukaryotes. In recent years, researchers in the field of metabolic engineering and synthetic biology have exploited various CRISPR-based pathway engineering approaches, which are both effective and convenient, as well as valuable from a theoretical standpoint. In this review, we systematically summarize novel pathway engineering techniques and strategies based on CRISPR nucleases system, CRISPR interference (CRISPRi), and CRISPR activation (CRISPRa), including figures and descriptions for easy understanding, with the aim to facilitate their broader application among fellow researchers.

高效的代谢途径是高效细胞工厂的关键。然而，工程化的同源或异源多基因途径可能不平衡、效率低下并导致潜在有毒中间体的积累。因此，必须以最佳方式构建路径，以最大限度地减少这些负面影响并最大限度地提高催化效率。随着 CRISPR 技术的发展，解决了之前通路工程和基因组编辑技术的一些问题，提供了更高的效率、更低的成本和易于定制的目标。此外，CRISPR 被证明在包括原核生物和真核生物在内的各种生物体中都具有强大和有效的作用。近年来，代谢工程和合成生物学领域的研究人员开发了各种基于 CRISPR 的通路工程方法，这既有效又方便，而且从理论的角度来看也很有价值。在这篇综述中，我们系统地总结了基于 CRISPR 核酸酶系统、CRISPR 干扰 (CRISPRi) 和 CRISPR 激活 (CRISPRa) 的新通路工程技术和策略，包括易于理解的图形和描述，旨在促进它们在同行中的更广泛应用。研究人员。