LAUSR

Microbial cell factories play a pivotal role in the field of biomanufacturing. Enhancing their construction efficiency and product yield is essential for advancing synthetic biology and its industrial implementation. In recent years, the rapid development of large DNA manipulation technologies provided powerful support for cloning, assembling, delivering, and rearranging large DNA. This review hence systematically summarizes the core principles and recent advances in four categories of large DNA manipulation techniques. It highlights their key roles in access to complex biosynthetic gene clusters, constructing multigene biosynthetic pathways, introducing complex genetic modules into microbial chassis, structural rewiring, and modular reconstruction for metabolic network optimization. Furthermore, this review explores the emerging trend of large DNA manipulation technologies to advance microbial cell factories. This review is expected to serve as a technical reference for advancing large DNA manipulation technologies and extending their applications toward the construction of high-performance microbial cell factories.