A history of genome editing in <scp><i>Saccharomyces cerevisiae</i></scp>

Alexander, William G.

doi:10.1002/yea.3300

Cited by 16 publications

(7 citation statements)

References 63 publications

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“…Although these technologies provide very useful insights, they are often difficult to implement and/or rely on the use of genetic markers. For this reason, the development of the CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated) system has boosted the field of genome engineering [25–27]. This system, initially derived from immune systems of bacteria, consists of an endonuclease encoded by the Cas9 gene of Streptococcus pyogenes and a short RNA that guides the endonuclease at the targeted genomic locus.…”

Section: Introductionmentioning

confidence: 99%

Reshuffling yeast chromosomes with CRISPR/Cas9

et al. 2019

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Genome engineering is a powerful approach to study how chromosomal architecture impacts phenotypes. However, quantifying the fitness impact of translocations independently from the confounding effect of base substitutions has so far remained challenging. We report a novel application of the CRISPR/Cas9 technology allowing to generate with high efficiency both uniquely targeted and multiple concomitant reciprocal translocations in the yeast genome. Targeted translocations are constructed by inducing two double-strand breaks on different chromosomes and forcing the trans-chromosomal repair through homologous recombination by chimerical donor DNAs. Multiple translocations are generated from the induction of several DSBs in LTR repeated sequences and promoting repair using endogenous uncut LTR copies as template. All engineered translocations are markerless and scarless. Targeted translocations are produced at base pair resolution and can be sequentially generated one after the other. Multiple translocations result in a large diversity of karyotypes and are associated in many instances with the formation of unanticipated segmental duplications. To test the phenotypic impact of translocations, we first recapitulated in a lab strain the SSU1/ECM34 translocation providing increased sulphite resistance to wine isolates. Surprisingly, the same translocation in a laboratory strain resulted in decreased sulphite resistance. However, adding the repeated sequences that are present in the SSU1 promoter of the resistant wine strain induced sulphite resistance in the lab strain, yet to a lower level than that of the wine isolate, implying that additional polymorphisms also contribute to the phenotype. These findings illustrate the advantage brought by our technique to untangle the phenotypic impacts of structural variations from confounding effects of base substitutions. Secondly, we showed that strains with multiple translocations, even those devoid of unanticipated segmental duplications, display large phenotypic diversity in a wide range of environmental conditions, showing that simply reconfiguring chromosome architecture is sufficient to provide fitness advantages in stressful growth conditions.

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Section: Introductionmentioning

confidence: 99%

Reshuffling yeast chromosomes with CRISPR/Cas9

et al. 2019

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“… 5 Walker and Hill ( 2016 ). 6 Botstein et al ( 1997 ), Legras et al ( 2007 ), Liti ( 2015 ), Bilinski et al ( 2017 ), and Alexander ( 2018 ). 7 Walker ( 2009 ), Wang et al ( 2012 ), Goddard and Greig ( 2015 ), Ramazzotti et al ( 2019 ), and Meriggi et al ( 2020 ).…”

Section: Variety Of Yeast Species and Their Application In Alcoholic ...mentioning

confidence: 99%

The potential for Scotch Malt Whisky flavour diversification by yeast

Daute,

Jack,

Walker

2024

FEMS Yeast Research

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Scotch Whisky, a product of high importance to Scotland, has gained global approval for its distinctive qualities derived from the traditional production process which is defined in law. However, ongoing research continuously enhances Scotch Whisky production and is fostering a diversification of flavour profiles. To be classified as Scotch Whisky, the final spirit needs to retain the aroma and taste of “Scotch”. While each production step contributes significantly to whisky flavour—from malt preparation and mashing to fermentation, distillation, and maturation—the impact of yeast during fermentation is crucially important. Not only does the yeast convert the sugar to alcohol, it also produces important volatile compounds, for example esters and higher alcohols, that contribute to the final flavour profile of whisky. The yeast chosen for whisky fermentations can significantly influence whisky flavour, so the yeast strain employed is of high importance. This review explores the role of yeast in Scotch Whisky production and its influence on flavour diversification. Furthermore, an extensive examination of non-conventional yeasts employed in brewing and winemaking is undertaken to assess their potential suitability for adoption as Scotch Whisky yeast strains, followed by a review of methods for evaluating new yeast strains.

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“…Among them, for example, biopharmaceutical recombinant peptide hormone, insulin, has been produced by genetically engineered S. cerevisiae strains for many years. Many pharmaceutical companies have chosen this yeast as the most suited host organism to produce a large variety of recombinant products due to its well-known genetics, physiology, biochemistry, and genetic engineering background, the availability of genetic tools, and the suitability of dense and large scale fermentation [16][17][18].…”

Section: Introductionmentioning

confidence: 99%

Fermentation Strategies for Production of Pharmaceutical Terpenoids in Engineered Yeast

2021

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Terpenoids, also known as isoprenoids, are a broad and diverse class of plant natural products with significant industrial and pharmaceutical importance. Many of these natural products have antitumor, anti-inflammatory, antibacterial, antiviral, and antimalarial effects, support transdermal absorption, prevent and treat cardiovascular diseases, and have hypoglycemic activities. Production of these compounds are generally carried out through extraction from their natural sources or chemical synthesis. However, these processes are generally unsustainable, produce low yield, and result in wasting of substantial resources, most of them limited. Microbial production of terpenoids provides a sustainable and environment-friendly alternative. In recent years, the yeast Saccharomyces cerevisiae has become a suitable cell factory for industrial terpenoid biosynthesis due to developments in omics studies (genomics, transcriptomics, metabolomics, proteomics), and mathematical modeling. Besides that, fermentation development has a significant importance on achieving high titer, yield, and productivity (TYP) of these compounds. Up to now, there have been many studies and reviews reporting metabolic strategies for terpene biosynthesis. However, fermentation strategies have not been yet comprehensively discussed in the literature. This review summarizes recent studies of recombinant production of pharmaceutically important terpenoids by engineered yeast, S. cerevisiae, with special focus on fermentation strategies to increase TYP in order to meet industrial demands to feed the pharmaceutical market. Factors affecting recombinant terpenoids production are reviewed (strain design and fermentation parameters) and types of fermentation process (batch, fed-batch, and continuous) are discussed.

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A history of genome editing in Saccharomyces cerevisiae

Cited by 16 publications

References 63 publications

Reshuffling yeast chromosomes with CRISPR/Cas9

Reshuffling yeast chromosomes with CRISPR/Cas9

The potential for Scotch Malt Whisky flavour diversification by yeast

Fermentation Strategies for Production of Pharmaceutical Terpenoids in Engineered Yeast

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