Metabolic engineering in budding yeast using CRISPR/dCas9

FUNDING: 9 weeks (full time, 37 hrs per week, £10.57 per hour, £690 consumables)
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Dr Sandesh Swamidatta, Biology, York

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The budding yeast, Saccharomyces cerevisiae, is a favoured microbial system for bio-based production of high value chemicals and natural products. The wide range of tools available for this species also makes it ideal for testing new enzymes and pathways via metabolic engineering. In this project, the student will test ways of increasing squalene accumulation, a known precursor to sterols and triterpenoids. This work connects with the ongoing work in the Lichman lab, to elucidate the biosynthetic pathway for triterpenoid alkaloids using S. cerevisiae as a model system. The student will employ CRISPR/dCas9 approach to repress the expression of a gene downstream of squalene and validate the success by measuring squalene levels using gas chromatography mass spectrometry (GC/MS). The student will be able to learn aspects of molecular cloning, yeast culturing and growth experiments, genome engineering, metabolite extractions and analysis using GCMS, and experimental data analysis and interpretation. The project is suitable for a student with strong interest in molecular biology, metabolic engineering or synthetic biology. The student will take part in regular lab meetings and journal clubs as part of Lichman lab. Students will be expected to present their findings orally at a research day in York in September 2023.