CRISPR–Cas system enables fast and simple genome editing of industrial Saccharomyces cerevisiae strains

Research output: Research - peer-reviewJournal article – Annual report year: 2015

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@article{fbbdd9c057e14ab59a23739654596b06,
title = "CRISPR–Cas system enables fast and simple genome editing of industrial Saccharomyces cerevisiae strains",
abstract = "There is a demand to develop 3rd generation biorefineries that integrate energy production with the production of higher value chemicals from renewable feedstocks. Here, robust and stress-tolerant industrial strains of Saccharomyces cerevisiae will be suitable production organisms. However, their genetic manipulation is challenging, as they are usually diploid or polyploid. Therefore, there is a need to develop more efficient genetic engineering tools. We applied a CRISPR–Cas9 system for genome editing of different industrial strains, and show simultaneous disruption of two alleles of a gene in several unrelated strains with the efficiency ranging between 65{\%} and 78{\%}. We also achieved simultaneous disruption and knock-in of a reporter gene, and demonstrate the applicability of the method by designing lactic acid-producing strains in a single transformation event, where insertion of a heterologous gene and disruption of two endogenous genes occurred simultaneously. Our study provides a foundation for efficient engineering of industrial yeast cell factories.",
keywords = "CRISPR–Cas9, Genome editing, Industrial yeast, Biorefineries, Chemical production",
author = "Vratislav Stovicek and Irina Borodina and Jochen F{\"o}rster",
note = "This is an open access article under the CCBY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).",
year = "2015",
doi = "10.1016/j.meteno.2015.03.001",
language = "English",
volume = "2",
pages = "13--22",
journal = "Metabolic Engineering Communications",
issn = "2214-0301",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - CRISPR–Cas system enables fast and simple genome editing of industrial Saccharomyces cerevisiae strains

AU - Stovicek,Vratislav

AU - Borodina,Irina

AU - Förster,Jochen

N1 - This is an open access article under the CCBY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PY - 2015

Y1 - 2015

N2 - There is a demand to develop 3rd generation biorefineries that integrate energy production with the production of higher value chemicals from renewable feedstocks. Here, robust and stress-tolerant industrial strains of Saccharomyces cerevisiae will be suitable production organisms. However, their genetic manipulation is challenging, as they are usually diploid or polyploid. Therefore, there is a need to develop more efficient genetic engineering tools. We applied a CRISPR–Cas9 system for genome editing of different industrial strains, and show simultaneous disruption of two alleles of a gene in several unrelated strains with the efficiency ranging between 65% and 78%. We also achieved simultaneous disruption and knock-in of a reporter gene, and demonstrate the applicability of the method by designing lactic acid-producing strains in a single transformation event, where insertion of a heterologous gene and disruption of two endogenous genes occurred simultaneously. Our study provides a foundation for efficient engineering of industrial yeast cell factories.

AB - There is a demand to develop 3rd generation biorefineries that integrate energy production with the production of higher value chemicals from renewable feedstocks. Here, robust and stress-tolerant industrial strains of Saccharomyces cerevisiae will be suitable production organisms. However, their genetic manipulation is challenging, as they are usually diploid or polyploid. Therefore, there is a need to develop more efficient genetic engineering tools. We applied a CRISPR–Cas9 system for genome editing of different industrial strains, and show simultaneous disruption of two alleles of a gene in several unrelated strains with the efficiency ranging between 65% and 78%. We also achieved simultaneous disruption and knock-in of a reporter gene, and demonstrate the applicability of the method by designing lactic acid-producing strains in a single transformation event, where insertion of a heterologous gene and disruption of two endogenous genes occurred simultaneously. Our study provides a foundation for efficient engineering of industrial yeast cell factories.

KW - CRISPR–Cas9

KW - Genome editing

KW - Industrial yeast

KW - Biorefineries

KW - Chemical production

U2 - 10.1016/j.meteno.2015.03.001

DO - 10.1016/j.meteno.2015.03.001

M3 - Journal article

VL - 2

SP - 13

EP - 22

JO - Metabolic Engineering Communications

T2 - Metabolic Engineering Communications

JF - Metabolic Engineering Communications

SN - 2214-0301

ER -