Construction and Validation of a CRISPR/dCpf1-Based Transcriptional Regulatory System in Saccharomyces Cerevisiae

Qing Feng; Jun Li

doi:10.3233/SHTI230900

Construction and Validation of a CRISPR/dCpf1-Based Transcriptional Regulatory System in Saccharomyces Cerevisiae

Stud Health Technol Inform. 2023 Nov 23:308:680-688. doi: 10.3233/SHTI230900.

Authors

Qing Feng¹, Jun Li¹

Affiliation

¹ Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering, Ministry of Industry and Information Technology, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 102488, China.

PMID: 38007799
DOI: 10.3233/SHTI230900

Abstract

Saccharomyces cerevisiae has been extensively studied and applied as a model microbial platform for efficient and sustainable production of biofuels, chemicals and natural products. In recent years, the application of CRISPR system in genome editing and transcriptional regulation has greatly improved the efficiency of microbial genome editing. To achieve transcriptional regulation the CRISPR/dCpf1-mediated CRISPRa/i transcriptional regulation system was constructed in this study. By constructing crRNA arrays and adding different activation proteins, repression efficiencies from 62% to 177% for the CRISPRi system and activation efficiencies from 154% to 320% for the CRISPRa system were achieved in the eGFP gene reporter system.

Keywords: CRISPR-dCpf1; CRISPRa/i; Saccharomyces cerevisiae.

MeSH terms

CRISPR-Cas Systems* / genetics
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Gene Editing
Gene Expression Regulation
Saccharomyces cerevisiae* / genetics
Saccharomyces cerevisiae* / metabolism