Chassis provides a setting for the expression of heterologous pathway genes, which often requires extensive engineering to achieve complete functions. Traditionally, chassis engineering relies on gene deletion/overexpression for the regulation of precursor/cofactor supply and product transportation, which has generated thousands of high-performance strains. With the development of synthetic biology, chassis modifications have expanded to the synthesis of artificial cellular machineries, creating synthetic cells for the biosynthesis of bioproducts. In this review, we will discuss the development of chassis engineering technologies, termed the first-generation and second-generation technologies, and their applications in the creation of chassis for the production of valued-added chemicals, with an emphasis on synthetic chassis and their applications and potential. The development of chassis engineering technologies will advance rational design and construction of customized chassis for the manufacturing of target bioproducts.
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