FOOD SCIENCE ›› 2019, Vol. 40 ›› Issue (6): 151-158.doi: 10.7506/spkx1002-6630-20180125-317

• Bioengineering • Previous Articles     Next Articles

Gene Deletion in Saccharomyces cerevisiae Using CRISPR-Cas9 System

NIU Xiaodi1, LI Tianming1, LIU Jinlei1, YANG Tianyong1, LI Ziyi2, FENG Huiyong1,*   

  1. 1. College of Biological Science and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, China; 2. College of Arts and Sciences, College Park, University of Maryland, Maryland 20742, USA
  • Online:2019-03-25 Published:2019-04-02

Abstract: In this study, CRISPR-Cas9-mediated gene knockout in diploid cells of Saccharomyces cerevisiae was investigated. The effectiveness of the CRISPR-Cas9 system was tested using the can1 knockout phenotype. The experimental results showed that the inactivation efficiency of can1 gene reached 4%. The pdc, adh3, adh2, adh1 and pdh genes were knocked out by this system, respectively, and the efficiency of gene editing was 4/48, 3/48, 1/48, 3/28 and 1/16, respectively. This study also developed a procedure for continuous gene knockout, and it took 17 days to knock out pdc, adh3, and adh2 using this procedure. It took 6 days to simultaneously disrupt adh5 and lip with 9/32 and 10/32 efficiencies, respectively.

Key words: Saccharomyces cerevisiae, CRISPR-Cas9, genome engineering, gene knockout

CLC Number: