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Data from Development of a CRISPR/Cas9 System for High-Efficiency Multiplexed Gene Deletion in Rhodosporidium toruloides

Dataset ·
 [1];  [1];  [2]
  1. Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL
  2. Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL; Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL; Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL; Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
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The oleaginous yeast Rhodosporidium toruloides is considered a promising candidate for production of chemicals and biofuels thanks to its ability to grow on lignocellulosic biomass, and its high production of lipids and carotenoids. However, efforts to engineer this organism are hindered by a lack of suitable genetic tools. Here we report the development of a CRISPR/Cas9 system for genome editing in R. toruloides based on a fusion 5S rRNA–tRNA promoter for guide RNA (gRNA) expression, capable of greater than 95% gene knockout for various genetic targets. Additionally, multiplexed double‐gene knockout mutants were obtained using this method with an efficiency of 78%. This tool can be used to accelerate future metabolic engineering work in this yeast.
Research Organization:
Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign
Sponsoring Organization:
U.S. Department of Energy (DOE)
DOE Contract Number:
SC0018420
OSTI ID:
3013653
Country of Publication:
United States
Language:
English

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Related Subjects

Conversion
Genome Engineering
Genomics
Transcriptomics