Cyanamide-inducible expression of homing nuclease I- Sce I for selectable marker removal and promoter characterisation in Saccharomyces cerevisiae .

Autor:	McDonnell L; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia., Evans S; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia., Lu Z; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia.; Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, 4072, Australia., Suchoronczak M; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Leighton J; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Ordeniza E; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Ritchie B; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Valado N; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Walsh N; School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Antoney J; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia., Wang C; College of Life Sciences, Shandong Agricultural University, Taian, Shandong Province, 271018, People's Republic of China., Luna-Flores CH; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia., Scott C; CSIRO Environment, Black Mountain Science and Innovation Park, Canberra, ACT, 2601, Australia., Speight R; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia.; Advanced Engineering Biology Future Science Platform, Commonwealth Scientific and Industrial Research Organisation (CSIRO), Black Mountain, ACT, 2601, Australia., Vickers CE; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia., Peng B; Centre of Agriculture and the Bioeconomy, School of Biology and Environmental Science, Faculty of Science, Queensland University of Technology, Brisbane, QLD, 4000, Australia.; ARC Centre of Excellence in Synthetic Biology, Australia.; Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, 4072, Australia.
Jazyk:	angličtina
Zdroj:	Synthetic and systems biotechnology [Synth Syst Biotechnol] 2024 Jun 28; Vol. 9 (4), pp. 820-827. Date of Electronic Publication: 2024 Jun 28 (Print Publication: 2024).
DOI:	10.1016/j.synbio.2024.06.009
Abstrakt:	In synthetic biology, microbial chassis including yeast Saccharomyces cerevisiae are iteratively engineered with increasing complexity and scale. Wet-lab genetic engineering tools are developed and optimised to facilitate strain construction but are often incompatible with each other due to shared regulatory elements, such as the galactose-inducible ( GAL ) promoter in S. cerevisiae . Here, we prototyped the cyanamide-induced I- Sce I expression, which triggered double-strand DNA breaks (DSBs) for selectable marker removal. We further combined cyanamide-induced I- Sce I-mediated DSB and maltose-induced MazF-mediated negative selection for plasmid-free in situ promoter substitution, which simplified the molecular cloning procedure for promoter characterisation. We then characterised three tetracycline-inducible promoters showing differential strength, a non-leaky β-estradiol-inducible promoter, cyanamide-inducible DDI2 promoter, bidirectional MAL32/MAL31 promoters, and five pairs of bidirectional GAL1/GAL10 promoters. Overall, alternative regulatory controls for genome engineering tools can be developed to facilitate genomic engineering for synthetic biology and metabolic engineering applications. Competing Interests: Authors declare no conflicts of interest. (© 2024 The Authors.)
Databáze:	MEDLINE
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