文章基本信息

标题：Type I-F CRISPR-PAIR platform for multi-mode regulation to boost extracellular electron transfer in Shewanella oneidensis
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作者：Yaru Chen ; Meijie Cheng ; Hao Song 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2022
卷号：25
期号：6
页码：1-17
DOI：10.1016/j.isci.2022.104491
语种：English
出版社：Elsevier
摘要：SummaryBio-electrochemical systems are based on extracellular electron transfer (EET), whose efficiency relates to the expression level of numerous genes. However, the lack of multi-functional tools for gene activation and repression hampers the enhancement of EET in electroactive microorganisms (EAMs). We thus develop a type I-FCRISPR/PaeCascade-RpoD-mediatedactivation andinhibitionregulation (CRISPR-PAIR) platform in the model EAM,Shewanella oneidensisMR-1. Gene activation is achieved (3.8-fold) through fusing activator RpoD (σ70) to Cas7 when targeting the prioritized loci upstream of the transcription start site. Gene inhibition almost has no position preference when targeting the open reading frame, which makes the design of crRNAs easy and flexible. Then CRISPR-PAIR platform is applied to up-/down-regulate the expression of six endogenous genes, resulting in the improved EET efficiency. Moreover, simultaneous gene activation and inhibition are achieved inS. oneidensisMR-1. CRISPR-PAIR platform offers a programmable methodology for dual regulation, facilitating in-depth EET studies inShewanellaspp.Graphical abstractDisplay OmittedHighlights•CRISPR-PAIR platform enables both gene activation and inhibition inShewanella oneidensis•An efficient type I-F CRISPR-Cas tool is developed forS. oneidensis•Transcription regulation of endogenous genes enhances extracellular electron transfer (EET)Biochemistry; Biochemical engineering; Metabolic engineering