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.
Stud Health Technol Inform. 2023 Nov 23;308:680-688. doi: 10.3233/SHTI230900.
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.
酿酒酵母作为一种模式微生物平台，已被广泛研究和应用，用于高效和可持续地生产生物燃料、化学品和天然产物。近年来，CRISPR 系统在基因组编辑和转录调控中的应用极大地提高了微生物基因组编辑的效率。为了实现转录调控，本研究构建了 CRISPR/dCpf1 介导的 CRISPRa/i 转录调控系统。通过构建 crRNA 阵列并添加不同的激活蛋白，在 eGFP 基因报告系统中，CRISPRi 系统的抑制效率从 62%提高到 177%，CRISPRa 系统的激活效率从 154%提高到 320%。
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