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Multiplex Genome Engineering of Polyploid Industrial Yeast Strains Using an Optimized CRISPR/Cas9System
Published: February 20, 2018
Posted: August 02, 2021

Objective

Develop an efficient CRISPR/Cas9 system for industrial yeast strain engineering.

Approach

  • Increasing gRNA abundance via super-high copy number plasmids.

Results/Impact

  • Plasmids with higher copy numbers increased gRNA levels, resulting in higher genome editing efficiencies (up to 100% efficiency).
    • Up to 12 alleles were disrupted in a single step with 100% knock-out efficiency.
    • Facile engineering of industrial yeast strains for practical applications (i.e., biofuel production).
  • Optimized CRISPR/Cas9 system provides a powerful tool for the development of industrial yeast-based microbial cell factories.

Lian, J. et al. “Engineered CRISPR/Cas9 system for multiplex genome engineering of polyploid industrial yeast strains.” Biotechnology and Bioengineering 115(6), 1630–1635 (2017). [DOI:10.1002/bit.26569]

Topic Areas:

  • Research Area: Genomic Analysis and Systems Biology
  • Research Area: Microbes and Communities
  • Research Area: Biosystems Design

Division: SC-33.2 Biological Systems Science Division, BER

 

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