Monthly Meeting of SB Club this March 25, 2024

Monthly Meeting of SB Club this March 25, 2024

We would like to inform you that the monthly online meeting of Synthetic Biology Club (SB club) will be held this Monday, March 25, 2024 at 12h00 (EST). Please, Click here to register after which you will receive the meeting link (zoom).

Title: A journey in the optimisation of CRISPR-Cas9-based genome editing tools for Yarrowia lipolytica

Presenter: Ahmad Saleh, Associate Professor at Laval University, Principal Investigator at the Abdel-Mawgoud Synthetic Biology Lab and Director of PROTEO-ULaval Research Center

Time: Monday, March 25, 2024 at 12h00 (EST).

Place: Online (via Zoom). The zoom link of the meeting is provided by email after your registration. Click here to register.

Abstract: Yarrowia lipolytica is an important oleaginous yeast used in the production of specialty chemicals and has a great potential for further applications in lipid biotechnology. Harnessing the full potential of Y. lipolytica is, however, limited by its inherent recalcitrance to traditional genetic manipulations. Current CRISPR/Cas9-based methods resulted in great advances in genome editing of this yeast, yet it had limitations in terms of the number of editable genomic targets as well as in the efficiency at which they can be edited. We further improved CRISPR-Cas9 editing protocol by optimising the expression cassettes of gRNA and Cas9. For optimisation of gRNA expression, we used tools of secondary RNA structure prediction that guided our redesign of the tRNA-sgRNA fusions used for the expression of single guide RNA (sgRNA). This resulted in high efficiency CRISPR/cas9 genome editing at chromosomal loci that failed gene editing or were previously editable at very low efficiencies and using templates with short homology arms. For optimisation of Cas9 expression, we used tools of prediction of promoter elements that allowed the reengineering of a short TEF promoter 5-fold higher gene expression than currently engineered TEF promoters. Expression of Cas9 under the reengineered short TEF promoter increased gene integration efficiency. These optimized genome editing tools are supposed to enhance genetic and metabolic engineering the industrial yeast, Yarrowia lipolytica.

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