Multiplex Genome Engineering of Polyploid Industrial Yeast Strains Using an Optimized CRISPR/Cas9System

The Science

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

The 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.


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

Principal Investigator(s)

Huimin Zhao
University of Illinois at Urbana-Champaign


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]