The synthetic biology toolkit for baker’s yeast,Saccharomyces cerevisiae,includes extensive genome engineering toolkits and parts repositories.However,with the increasing complexity of engineering tasks and versatile...The synthetic biology toolkit for baker’s yeast,Saccharomyces cerevisiae,includes extensive genome engineering toolkits and parts repositories.However,with the increasing complexity of engineering tasks and versatile applications of this model eukaryote,there is a continued interest to expand and diversify the rational engineering capabilities in this chassis by FAIR(findable,accessible,interoperable,and reproducible)compliance.In this study,we designed and characterised 41 synthetic guide RNA sequences to expand the CRISPR-based genome engineering capabilities for easy and efficient replacement of genomically encoded elements.Moreover,we characterize in high temporal resolution 20 native promoters and 18 terminators using fluorescein and LUDOX CL-X as references for GFP expression and OD600 measurements,respectively.Additionally,all data and reported analysis is provided in a publicly accessible jupyter notebook providing a tool for researchers with low-coding skills to further explore the generated data as well as a template for researchers to write their own scripts.We expect the data,parts,and databases associated with this study to support a FAIR-compliant resource for further advancing the engineering of yeasts.展开更多
基金This study is supported by grants from the Novo Nordisk Foundation(NNF10CC1016517)Horizon 2020(MIAMi,814645)BioInnovation Institute Foundation(NNF19OC0055591 and NNF20SA0067054).
文摘The synthetic biology toolkit for baker’s yeast,Saccharomyces cerevisiae,includes extensive genome engineering toolkits and parts repositories.However,with the increasing complexity of engineering tasks and versatile applications of this model eukaryote,there is a continued interest to expand and diversify the rational engineering capabilities in this chassis by FAIR(findable,accessible,interoperable,and reproducible)compliance.In this study,we designed and characterised 41 synthetic guide RNA sequences to expand the CRISPR-based genome engineering capabilities for easy and efficient replacement of genomically encoded elements.Moreover,we characterize in high temporal resolution 20 native promoters and 18 terminators using fluorescein and LUDOX CL-X as references for GFP expression and OD600 measurements,respectively.Additionally,all data and reported analysis is provided in a publicly accessible jupyter notebook providing a tool for researchers with low-coding skills to further explore the generated data as well as a template for researchers to write their own scripts.We expect the data,parts,and databases associated with this study to support a FAIR-compliant resource for further advancing the engineering of yeasts.
