Designing strong inducible synthetic promoters in yeasts.

Autor: Tominaga M; Engineering Biology Research Center, Kobe University, Kobe, Japan.; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan., Shima Y; Engineering Biology Research Center, Kobe University, Kobe, Japan., Nozaki K; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan., Ito Y; Engineering Biology Research Center, Kobe University, Kobe, Japan.; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan., Someda M; Pharma Foods International Co. Ltd., Kyoto, Japan., Shoya Y; Pharma Foods International Co. Ltd., Kyoto, Japan., Hashii N; Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kawasaki, Kanagawa, Japan., Obata C; Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kawasaki, Kanagawa, Japan., Matsumoto-Kitano M; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan., Suematsu K; Engineering Biology Research Center, Kobe University, Kobe, Japan., Matsukawa T; Engineering Biology Research Center, Kobe University, Kobe, Japan., Hosoya K; Engineering Biology Research Center, Kobe University, Kobe, Japan., Hashiba N; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan., Kondo A; Engineering Biology Research Center, Kobe University, Kobe, Japan.; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.; Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, Kobe, Japan.; Center for Sustainable Resource Science, RIKEN, Yokohama, Japan., Ishii J; Engineering Biology Research Center, Kobe University, Kobe, Japan. junjun@port.kobe-u.ac.jp.; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan. junjun@port.kobe-u.ac.jp.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Dec 19; Vol. 15 (1), pp. 10653. Date of Electronic Publication: 2024 Dec 19.
DOI: 10.1038/s41467-024-54865-z
Abstrakt: Inducible promoters are essential for precise control of target gene expression in synthetic biological systems. However, engineering eukaryotic promoters is often more challenging than engineering prokaryotic promoters due to their greater mechanistic complexity. In this study, we describe a simple and reliable approach for constructing strongly inducible synthetic promoters with minimum leakiness in yeasts. The results indicate that the leakiness of yeast-inducible synthetic promoters is primarily the result of cryptic transcriptional activation of heterologous sequences that may be avoided by appropriate insulation and operator mutagenesis. Our promoter design approach has successfully generated robust, inducible promoters that achieve a > 10 3 -fold induction in reporter gene expression. The utility of these promoters is demonstrated by using them to produce various biologics with titers up to 2 g/L, including antigens designed to raise specific antibodies against a SARS-CoV-2 omicron variant through chicken immunization.
Competing Interests: Competing interests: M.S. and Y.Shoya are employees of Pharma Foods International Co. Ltd. M.T., K.N., A.K., and J.I. are inventors of pending patent applications submitted by Kobe University that cover the inducible synthetic promoters described in this study (PCT/JP2021/044573, PCT/JP2021/044574, and JP2020202234). Y.I. is the inventor of one of the above pending patent applications (PCT/JP2021/044573). M.T., Y.I., M.M.-K., M.S., Y.Shoya., A.K., and J.I. are inventors of the pending patent application submitted by Kobe University and Pharma Foods International Co. Ltd. that covers the IgY antibody obtained in this study (JP2023188697). The other authors declare no competing interests.
(© 2024. The Author(s).)
Databáze: MEDLINE
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