@article{d3a6f03083ed417f8d0db6e9e39db53b,
title = "Advancing Metabolic Engineering of Saccharomyces cerevisiae Using the CRISPR/Cas System",
abstract = "Thanks to its ease of use, modularity, and scalability, the clustered regularly interspaced short palindromic repeats (CRISPR) system has been increasingly used in the design and engineering of Saccharomyces cerevisiae, one of the most popular hosts for industrial biotechnology. This review summarizes the recent development of this disruptive technology for metabolic engineering applications, including CRISPR-mediated gene knock-out and knock-in as well as transcriptional activation and interference. More importantly, multi-functional CRISPR systems that combine both gain- and loss-of-function modulations for combinatorial metabolic engineering are highlighted.",
keywords = "Saccharomyces cerevisiae, gene regulation, genome engineering, metabolic engineering, synthetic biology",
author = "Jiazhang Lian and Mohammad HamediRad and Huimin Zhao",
note = "Funding Information: API, application program interface; BDO, 2,3-butanediol; CDS, coding sequences; CHAnGE, CRISPR/Cas9 and homology-directed repair assisted genome-scale engineering; CRISPR, clustered regularly interspaced short palindromic repeats; CRISPRa, CRISPR activation; CRISPRd, CRISPR mediated gene deletion; CRISPRi, CRISPR interference; CRISPR-AID, CRISPRa, CRIDPRi, and CRISPRd; dsOligo, double-strand oligonucleotides; GRAS, generally recognized as safe; gRNA, guide RNA; HDR, homology directed repair; HDV, hepatitis delta virus; HI-CRISPR, homology integrated CRISPR/Cas system; NLS, nuclear localization signal; PAM, protospacer adjacent motif; PDH, pyruvate dehydrogenase; Pol II (III), RNA polymerase (II) III; RGR, ribozyme-gRNA-ribozyme; scRNA, scaffold RNA; STEPS, systematically test enzyme perturbation sensitivities; VPR, VP64-p65-Rta (tripartite activator). This work was supported by the U.S. Department of Energy (DE-SC0018260 and DE-SC0018420). J.L. also acknowledges the support of the Shen Postdoc Fellowship from the University of Illinois at Urbana-Champaign and the Fundamental Research Funds for the Central Universities and Startup Fund from Zhejiang University.",
year = "2018",
month = sep,
doi = "10.1002/biot.201700601",
language = "English (US)",
volume = "13",
journal = "Biotechnology Journal",
issn = "1860-6768",
publisher = "Wiley-VCH Verlag",
number = "9",
}