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• W2975176095 abstract "ABSTRACT We present a teaching protocol suitable for demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of industrially relevant yeasts Saccharomyces cerevisiae and Yarrowia lipolytica, using β-carotene production as a case study. The protocol details all steps required to generate DNA parts, transform and genotype yeast, and perform a phenotypic screen to determine β-carotene production. The protocol is intended to be used as an instruction manual for a two-week practical course aimed at MSc and PhD students. The protocol details all necessary steps for students to engineer yeast to produce β-carotene and serves as a practical introduction to the principles of metabolic engineering including the concepts of boosting native precursor supply and alleviating rate-limiting steps. It also highlights key differences in the metabolism and heterologous production capacity of two industrially relevant yeast species. The protocol is divided into daily experiments covering a two week period and provides detailed instructions for every step meaning this protocol can be used ‘as is’ for a teaching course or as a case study for how yeast can be engineered to produce value-added molecules." @default.
• W2975176095 created "2019-10-03" @default.
• W2975176095 creator A5040605607 @default.
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• W2975176095 date "2019-09-26" @default.
• W2975176095 modified "2023-09-27" @default.
• W2975176095 title "A teaching protocol demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae and Yarrowia lipolytica" @default.
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• W2975176095 doi "https://doi.org/10.1093/femsyr/foz062" @default.
• W2975176095 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8260333" @default.
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