ArticleFEMS yeast research2019
A toolkit for rapid CRISPR-SpCas9 assisted construction of hexose-transport-deficient Saccharomyces cerevisiae strains.
Article in FEMS yeast research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed, 34 citations in OpenAlex.
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- Machine learning and comparative genomics approaches for the discovery of xylose transporters in yeast.Biotechnology for biofuels and bioproducts · 2022Article
- Engineering proton-coupled hexose uptake in Saccharomyces cerevisiae for improved ethanol yield.Biotechnology for biofuels and bioproducts · 2022Article
- Strategies for Efficient Expression of Heterologous Monosaccharide Transporters inJournal of fungi (Basel, Switzerland) · 2022Article
- Identification of novel pentose transporters in Kluyveromyces marxianus using a new screening platform.FEMS yeast research · 2021Article
- ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production.Biotechnology for biofuels · 2020Article
- Genome-scale metabolic rewiring improves titers rates and yields of the non-native product indigoidine at scale.Nature communications · 2020Article
- The CRISPR toolbox in medical mycology: State of the art and perspectives.PLoS pathogens · 2020Review
- Multiplex genome editing of microorganisms using CRISPR-Cas.FEMS microbiology letters · 2019Review
- Allele-specific genome editing using CRISPR-Cas9 is associated with loss of heterozygosity in diploid yeast.Nucleic acids research · 2019Article
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Authors and funding
10 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Hexose transporter-deficient yeast strains are valuable testbeds for the study of sugar transport by native and heterologous transporters. In the popular Saccharomyces cerevisiae strain EBY.VW4000, deletion of 21 transporters completely abolished hexose transport. However, repeated use of the LoxP/Cre system in successive deletion rounds also resulted in major chromosomal rearrangements, gene loss and phenotypic changes. In the present study, CRISPR/SpCas9 was used to delete the 21 hexose transporters in an S. cerevisiae strain from the CEN.PK family in only three deletion rounds, using 11 unique guide RNAs. Even upon prolonged cultivation, the resulting strain IMX1812 (CRISPR-Hxt0) was unable to consume glucose, while its growth rate on maltose was the same as that of a strain equipped with a full set of hexose transporters. Karyotyping and whole-genome sequencing of the CRISPR-Hxt0 strain with Illumina and Oxford Nanopore technologies did not reveal chromosomal rearrangements or other unintended mutations besides a few SNPs. This study provides a new, 'genetically unaltered' hexose transporter-deficient strain and supplies a CRISPR toolkit for removing all hexose transporter genes from most S. cerevisiae laboratory strains in only three transformation rounds.
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