Data from Development of a CRISPR/Cas9-Based Tool for Gene Deletion in Issatchenkia orientalis

Tran, Vinh; Cao, Mingfeng; Fatma, Zia; Song, Xiaofei; Zhao, Huimin

doi:10.13012/B2IDB-4661029_V1

Title: Data from Development of a CRISPR/Cas9-Based Tool for Gene Deletion in Issatchenkia orientalis

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Abstract

The nonconventional yeast Issatchenkia orientalis has emerged as a potential platform microorganism for production of organic acids due to its ability to grow robustly under highly acidic conditions. However, lack of efficient genetic tools remains a major bottleneck in metabolic engineering of this organism. Here we report that the autonomously replicating sequence (ARS) from Saccharomyces cerevisiae (ScARS) was functional for plasmid replication in I. orientalis, and the resulting episomal plasmid enabled efficient genome editing by the CRISPR/Cas9 system. The optimized CRISPR/Cas9-based system employed a fusion RPR1′-tRNA promoter for single guide RNA (sgRNA) expression and could attain greater than 97% gene disruption efficiency for various gene targets. Additionally, we demonstrated multiplexed gene deletion with disruption efficiencies of 90% and 47% for double gene and triple gene knockouts, respectively. This genome editing tool can be used for rapid strain development and metabolic engineering of this organism for production of biofuels and chemicals.

Authors:

;

; Fatma, Zia; Song, Xiaofei;

Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA; Department of Microbiology, Nankai University, Tianjin, China
Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA; Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA; Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA; Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

Publication Date:: Wed Jun 26 00:00:00 UTC 2019

DOE Contract Number:: SC0018420

Research Org.:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign

Sponsoring Org.:: U.S. Department of Energy (DOE)

Subject:: Conversion; Genome Engineering; Genomics; Transcriptomics

OSTI Identifier:: 3013685

DOI:: https://doi.org/10.13012/B2IDB-4661029_V1

Citation Formats


                    Tran, Vinh, Cao, Mingfeng, Fatma, Zia, Song, Xiaofei, and Zhao, Huimin. Data from Development of a CRISPR/Cas9-Based Tool for Gene Deletion in Issatchenkia orientalis.  United States: N. p., 2019. 
        Web.  doi:10.13012/B2IDB-4661029_V1.

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                    Tran, Vinh, Cao, Mingfeng, Fatma, Zia, Song, Xiaofei, & Zhao, Huimin. Data from Development of a CRISPR/Cas9-Based Tool for Gene Deletion in Issatchenkia orientalis.  United States.  doi:https://doi.org/10.13012/B2IDB-4661029_V1

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                    Tran, Vinh, Cao, Mingfeng, Fatma, Zia, Song, Xiaofei, and Zhao, Huimin. 2019.  
"Data from Development of a CRISPR/Cas9-Based Tool for Gene Deletion in Issatchenkia orientalis".  United States.  doi:https://doi.org/10.13012/B2IDB-4661029_V1.  https://www.osti.gov/servlets/purl/3013685. Pub date:Wed Jun 26 00:00:00 UTC 2019

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@article{osti_3013685,

  title        = {Data from Development of a CRISPR/Cas9-Based Tool for Gene Deletion in Issatchenkia orientalis},

  author       = {Tran, Vinh and Cao, Mingfeng and Fatma, Zia and Song, Xiaofei and Zhao, Huimin},

  abstractNote = {The nonconventional yeast Issatchenkia orientalis has emerged as a potential platform microorganism for production of organic acids due to its ability to grow robustly under highly acidic conditions. However, lack of efficient genetic tools remains a major bottleneck in metabolic engineering of this organism. Here we report that the autonomously replicating sequence (ARS) from Saccharomyces cerevisiae (ScARS) was functional for plasmid replication in I. orientalis, and the resulting episomal plasmid enabled efficient genome editing by the CRISPR/Cas9 system. The optimized CRISPR/Cas9-based system employed a fusion RPR1′-tRNA promoter for single guide RNA (sgRNA) expression and could attain greater than 97% gene disruption efficiency for various gene targets. Additionally, we demonstrated multiplexed gene deletion with disruption efficiencies of 90% and 47% for double gene and triple gene knockouts, respectively. This genome editing tool can be used for rapid strain development and metabolic engineering of this organism for production of biofuels and chemicals.},

  doi          = {10.13012/B2IDB-4661029_V1},

  journal      = {},

  number       = ,

  volume       = ,

  place        = {United States},

  year         = {Wed Jun 26 00:00:00 UTC 2019},

  month        = {Wed Jun 26 00:00:00 UTC 2019}

}

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DOI: https://doi.org/10.13012/B2IDB-4661029_V1

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