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CRISPRi screens reveal genes modulating yeast growth in lignocellulose hydrolysate

By Friederike Gutmann, Cosimo Jann, Filipa Pereira, Andreas Johansson, Lars Steinmetz, Kiran Raosaheb Patil

Posted 17 Sep 2020
bioRxiv DOI: 10.1101/2020.09.17.301416

Background Baker’s yeast is a widely used eukaryotic cell factory, producing a diverse range of compounds including biofuels and fine chemicals. The use of lignocellulose as feedstock offers the opportunity to run these processes in an environmentally sustainable way. However, the required hydrolysis pretreatment of lignocellulosic material releases toxic compounds that hamper yeast growth and consequently productivity. Results Here, we employ CRISPR interference in S. cerevisiae to identify genes modulating fermentative growth in plant hydrolysate and in presence of lignocellulosic toxins. We find that at least one third of hydrolysate-associated gene functions are explained by effects of known toxic compounds, such as the decreased growth of YAP1 or HAA1 , or increased growth of DOT6 knock-down strains in hydrolysate. Conclusion Our study confirms previously known genetic elements and uncovers new targets towards designing more robust yeast strains for the utilization of lignocellulose hydrolysate as sustainable feedstock, and, more broadly, paves the way for applying CRISPRi screens to improve industrial fermentation processes. ### Competing Interest Statement The authors have declared no competing interest. * ATc : Anhydrotetracycline CRISPRi : Clustered Regularly Interspaced Short Palindromic Repeats interference (d)Cas9 : (dead) Clustered Regularly Interspaced Short Palindromic Repeats-associated protein 9 HPLC : High performance liquid chromatography IC : Inhibitor compound cocktail MDS : Multidimensional scaling SCM : Synthetic complete medium

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