It is a routine task in metabolic engineering to introduce multicomponent pathways into a heterologous host for production of metabolites. However, this process sometimes may take weeks to months due to the lack of standardized genetic tools. Here, we present a method for the design and construction of biological parts based on the native genes and regulatory elements in Saccharomyces cerevisiae. We have developed highly efficient protocols (termed YeastFab Assembly) to synthesize these genetic elements as standardized biological parts, which can be used to assemble transcriptional units in a single-tube reaction. In addition, standardized characterization assays are developed using reporter constructs to calibrate the function of promoters.Furthermore, the assembled transcription units can be either assayed individually or applied to construct multigene metabolic pathways, which targets a genomic locus or a receiving plasmid effectively, through a simple in vitro reaction. Finally, using β-carotene biosynthesis pathway as an example, we demonstrate that our method allows us not only to construct and test a metabolic pathway in several days, but also to optimize the production through combinatorial assembly of a pathway using hundreds of regulatory biological parts.
Patrick Yizhi Cai (PC, Manchester) is Professor in Synthetic Genomics and a world-leading expert in synthetic chromosomes, with a highly interdisciplinary research group. In 2017, Prof. Cai’s team published 7 research articles in the journal of Science and featured on its cover. PC is the international coordinator for the (Sc2.0) Consortium, which is composed of over 10 top universities from 4 continents aiming to synthesize the world’s first synthetic eukaryotic genome. PC founded Edinburgh Genome Foundry, which is the largest automated DNA synthesis and assembly facility in academia today. PC regularly provides advice and consultancies to the Cabinet office, the Foreign Office and the Prime Minister’s Council for Science and Technologies. PC holds prestigious visiting professorships with MIT (US), MRC LMB at Cambridge (UK), Hong Kong University and Chinese Academy of Sciences (China). In 2022, PC was awarded a 5year EPSRC fellowship to work on biosecurity and biosafety mechanisms for synthetic genomes. In 2023, PC was awarded an ERC Consolidator Award to engineer non-coding RNAs using a synthetic genomics approach.
