Developing an endogenous quorum-sensing based CRISPRi circuit for autonomous and tunable dynamic regulation of multiple targets in Streptomyces.
Identifieur interne : 000129 ( Main/Exploration ); précédent : 000128; suivant : 000130Developing an endogenous quorum-sensing based CRISPRi circuit for autonomous and tunable dynamic regulation of multiple targets in Streptomyces.
Auteurs : Jinzhong Tian [République populaire de Chine] ; Gaohua Yang [République populaire de Chine] ; Yang Gu [République populaire de Chine] ; Xinqiang Sun [République populaire de Chine] ; Yinhua Lu [République populaire de Chine] ; Weihong Jiang [République populaire de Chine]Source :
- Nucleic acids research [ 1362-4962 ] ; 2020.
Descripteurs français
- KwdFr :
- MESH :
- génétique : Streptomyces.
- métabolisme : Sirolimus, Streptomyces.
- méthodes : Microbiologie industrielle.
- Détection du quorum, Régulation de l'expression des gènes bactériens, Systèmes CRISPR-Cas.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : Sirolimus.
- genetics : Streptomyces.
- metabolism : Streptomyces.
- methods : Industrial Microbiology.
- CRISPR-Cas Systems, Gene Expression Regulation, Bacterial, Quorum Sensing.
Abstract
Quorum-sensing (QS) mediated dynamic regulation has emerged as an effective strategy for optimizing product titers in microbes. However, these QS-based circuits are often created on heterologous systems and require careful tuning via a tedious testing/optimization process. This hampers their application in industrial microbes. Here, we design a novel QS circuit by directly integrating an endogenous QS system with CRISPRi (named EQCi) in the industrial rapamycin-producing strain Streptomyces rapamycinicus. EQCi combines the advantages of both the QS system and CRISPRi to enable tunable, autonomous, and dynamic regulation of multiple targets simultaneously. Using EQCi, we separately downregulate three key nodes in essential pathways to divert metabolic flux towards rapamycin biosynthesis and significantly increase its titers. Further application of EQCi to simultaneously regulate these three key nodes with fine-tuned repression strength boosts the rapamycin titer by ∼660%, achieving the highest reported titer (1836 ± 191 mg/l). Notably, compared to static engineering strategies, which result in growth arrest and suboptimal rapamycin titers, EQCi-based regulation substantially promotes rapamycin titers without affecting cell growth, indicating that it can achieve a trade-off between essential pathways and product synthesis. Collectively, this study provides a convenient and effective strategy for strain improvement and shows potential for application in other industrial microorganisms.
DOI: 10.1093/nar/gkaa602
PubMed: 32672817
PubMed Central: PMC7430639
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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wicri:level="1"><nlm:affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032</wicri:regionArea><wicri:noRegion>Shanghai 200032</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing 100039, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing 100039</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Yang, Gaohua" sort="Yang, Gaohua" uniqKey="Yang G" first="Gaohua" last="Yang">Gaohua Yang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032</wicri:regionArea><wicri:noRegion>Shanghai 200032</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing 100039, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing 100039</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Gu, Yang" sort="Gu, Yang" uniqKey="Gu Y" first="Yang" last="Gu">Yang Gu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032</wicri:regionArea><wicri:noRegion>Shanghai 200032</wicri:noRegion></affiliation></author><author><name sortKey="Sun, Xinqiang" sort="Sun, Xinqiang" uniqKey="Sun X" first="Xinqiang" last="Sun">Xinqiang Sun</name><affiliation wicri:level="1"><nlm:affiliation>XinChang Pharmaceutical Factory, Zhejiang medicine LTD, Xinchang 312500, Zhejiang Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>XinChang Pharmaceutical Factory, Zhejiang medicine LTD, Xinchang 312500, Zhejiang Province</wicri:regionArea><wicri:noRegion>Zhejiang Province</wicri:noRegion></affiliation></author><author><name sortKey="Lu, Yinhua" sort="Lu, Yinhua" uniqKey="Lu Y" first="Yinhua" last="Lu">Yinhua Lu</name><affiliation wicri:level="1"><nlm:affiliation>College of Life Sciences, Shanghai Normal University, Shanghai 200234, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences, Shanghai Normal University, Shanghai 200234</wicri:regionArea><wicri:noRegion>Shanghai 200234</wicri:noRegion></affiliation></author><author><name sortKey="Jiang, Weihong" sort="Jiang, Weihong" uniqKey="Jiang W" first="Weihong" last="Jiang">Weihong Jiang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032</wicri:regionArea><wicri:noRegion>Shanghai 200032</wicri:noRegion></affiliation></author></analytic><series><title level="j">Nucleic acids research</title><idno type="eISSN">1362-4962</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>CRISPR-Cas Systems (MeSH)</term><term>Gene Expression Regulation, Bacterial (MeSH)</term><term>Industrial Microbiology (methods)</term><term>Quorum Sensing (MeSH)</term><term>Sirolimus (metabolism)</term><term>Streptomyces (genetics)</term><term>Streptomyces (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Détection du quorum (MeSH)</term><term>Microbiologie industrielle (méthodes)</term><term>Régulation de l'expression des gènes bactériens (MeSH)</term><term>Sirolimus (métabolisme)</term><term>Streptomyces (génétique)</term><term>Streptomyces (métabolisme)</term><term>Systèmes CRISPR-Cas (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Sirolimus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Industrial Microbiology</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Sirolimus</term><term>Streptomyces</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Microbiologie industrielle</term></keywords><keywords scheme="MESH" xml:lang="en"><term>CRISPR-Cas Systems</term><term>Gene Expression Regulation, Bacterial</term><term>Quorum Sensing</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Détection du quorum</term><term>Régulation de l'expression des gènes bactériens</term><term>Systèmes CRISPR-Cas</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Quorum-sensing (QS) mediated dynamic regulation has emerged as an effective strategy for optimizing product titers in microbes. However, these QS-based circuits are often created on heterologous systems and require careful tuning via a tedious testing/optimization process. This hampers their application in industrial microbes. Here, we design a novel QS circuit by directly integrating an endogenous QS system with CRISPRi (named EQCi) in the industrial rapamycin-producing strain Streptomyces rapamycinicus. EQCi combines the advantages of both the QS system and CRISPRi to enable tunable, autonomous, and dynamic regulation of multiple targets simultaneously. Using EQCi, we separately downregulate three key nodes in essential pathways to divert metabolic flux towards rapamycin biosynthesis and significantly increase its titers. Further application of EQCi to simultaneously regulate these three key nodes with fine-tuned repression strength boosts the rapamycin titer by ∼660%, achieving the highest reported titer (1836 ± 191 mg/l). Notably, compared to static engineering strategies, which result in growth arrest and suboptimal rapamycin titers, EQCi-based regulation substantially promotes rapamycin titers without affecting cell growth, indicating that it can achieve a trade-off between essential pathways and product synthesis. Collectively, this study provides a convenient and effective strategy for strain improvement and shows potential for application in other industrial microorganisms.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">32672817</PMID><DateCompleted><Year>2020</Year><Month>11</Month><Day>09</Day></DateCompleted><DateRevised><Year>2020</Year><Month>11</Month><Day>09</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1362-4962</ISSN><JournalIssue CitedMedium="Internet"><Volume>48</Volume><Issue>14</Issue><PubDate><Year>2020</Year><Month>08</Month><Day>20</Day></PubDate></JournalIssue><Title>Nucleic acids research</Title><ISOAbbreviation>Nucleic Acids Res</ISOAbbreviation></Journal><ArticleTitle>Developing an endogenous quorum-sensing based CRISPRi circuit for autonomous and tunable dynamic regulation of multiple targets in Streptomyces.</ArticleTitle><Pagination><MedlinePgn>8188-8202</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1093/nar/gkaa602</ELocationID><Abstract><AbstractText>Quorum-sensing (QS) mediated dynamic regulation has emerged as an effective strategy for optimizing product titers in microbes. However, these QS-based circuits are often created on heterologous systems and require careful tuning via a tedious testing/optimization process. This hampers their application in industrial microbes. Here, we design a novel QS circuit by directly integrating an endogenous QS system with CRISPRi (named EQCi) in the industrial rapamycin-producing strain Streptomyces rapamycinicus. EQCi combines the advantages of both the QS system and CRISPRi to enable tunable, autonomous, and dynamic regulation of multiple targets simultaneously. Using EQCi, we separately downregulate three key nodes in essential pathways to divert metabolic flux towards rapamycin biosynthesis and significantly increase its titers. Further application of EQCi to simultaneously regulate these three key nodes with fine-tuned repression strength boosts the rapamycin titer by ∼660%, achieving the highest reported titer (1836 ± 191 mg/l). Notably, compared to static engineering strategies, which result in growth arrest and suboptimal rapamycin titers, EQCi-based regulation substantially promotes rapamycin titers without affecting cell growth, indicating that it can achieve a trade-off between essential pathways and product synthesis. Collectively, this study provides a convenient and effective strategy for strain improvement and shows potential for application in other industrial microorganisms.</AbstractText><CopyrightInformation>© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Tian</LastName><ForeName>Jinzhong</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing 100039, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Gaohua</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing 100039, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gu</LastName><ForeName>Yang</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sun</LastName><ForeName>Xinqiang</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>XinChang Pharmaceutical Factory, Zhejiang medicine LTD, Xinchang 312500, Zhejiang Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lu</LastName><ForeName>Yinhua</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>College of Life Sciences, Shanghai Normal University, Shanghai 200234, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Weihong</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences (CAS), Shanghai 200032, China.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Nucleic Acids Res</MedlineTA><NlmUniqueID>0411011</NlmUniqueID><ISSNLinking>0305-1048</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>W36ZG6FT64</RegistryNumber><NameOfSubstance UI="D020123">Sirolimus</NameOfSubstance></Chemical></ChemicalList><SupplMeshList><SupplMeshName Type="Organism" UI="C000640059">Streptomyces rapamycinicus</SupplMeshName></SupplMeshList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D064113" MajorTopicYN="Y">CRISPR-Cas Systems</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015964" MajorTopicYN="Y">Gene Expression Regulation, Bacterial</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007218" MajorTopicYN="N">Industrial Microbiology</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D053038" MajorTopicYN="Y">Quorum Sensing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020123" MajorTopicYN="N">Sirolimus</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013302" 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last="Tian">Jinzhong Tian</name></noRegion><name sortKey="Gu, Yang" sort="Gu, Yang" uniqKey="Gu Y" first="Yang" last="Gu">Yang Gu</name><name sortKey="Jiang, Weihong" sort="Jiang, Weihong" uniqKey="Jiang W" first="Weihong" last="Jiang">Weihong Jiang</name><name sortKey="Lu, Yinhua" sort="Lu, Yinhua" uniqKey="Lu Y" first="Yinhua" last="Lu">Yinhua Lu</name><name sortKey="Sun, Xinqiang" sort="Sun, Xinqiang" uniqKey="Sun X" first="Xinqiang" last="Sun">Xinqiang Sun</name><name sortKey="Tian, Jinzhong" sort="Tian, Jinzhong" uniqKey="Tian J" first="Jinzhong" last="Tian">Jinzhong Tian</name><name sortKey="Yang, Gaohua" sort="Yang, Gaohua" uniqKey="Yang G" first="Gaohua" last="Yang">Gaohua Yang</name><name sortKey="Yang, Gaohua" sort="Yang, Gaohua" uniqKey="Yang G" first="Gaohua" last="Yang">Gaohua Yang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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