Abstract
Traditional metabolic engineering faces numerous challenges in constructing microbial cell factories, such as inefficient gene editing, time-consuming and labor-intensive screening processes, and the complexity of multi-gene optimization. High-throughput (HTP) genome editing technology accelerates the optimization of microbial metabolic pathways by precisely and efficiently modifying multiple genes. Furthermore, HTP genome editing technology enables the rapid screening and modification of key enzymes or regulatory factors across multiple metabolic pathways, facilitating the analysis of complex regulatory mechanisms. These advantages make it a key enabling tool for both top-down analysis and bottom-up assembly of metabolic pathways. This review summarizes the mechanisms and applications of various HTP genome editing and discusses the development of HTP strategies that accelerate the design-build phase for microbial cell factories (MCFs). These advancements promised to significantly enhance the performance of MCFs and drive the next generation of sustainable, bio-based production technologies.