{"@context":{"@vocab":"https://cir.nii.ac.jp/schema/1.0/","rdfs":"http://www.w3.org/2000/01/rdf-schema#","dc":"http://purl.org/dc/elements/1.1/","dcterms":"http://purl.org/dc/terms/","foaf":"http://xmlns.com/foaf/0.1/","prism":"http://prismstandard.org/namespaces/basic/2.0/","cinii":"http://ci.nii.ac.jp/ns/1.0/","datacite":"https://schema.datacite.org/meta/kernel-4/","ndl":"http://ndl.go.jp/dcndl/terms/","jpcoar":"https://github.com/JPCOAR/schema/blob/master/2.0/"},"@id":"https://cir.nii.ac.jp/crid/1360021391864693888.json","@type":"Article","productIdentifier":[{"identifier":{"@type":"DOI","@value":"10.1111/mpp.13378"}},{"identifier":{"@type":"URI","@value":"https://bsppjournals.onlinelibrary.wiley.com/doi/pdf/10.1111/mpp.13378"}},{"identifier":{"@type":"DOI","@value":"10.1101/2023.04.20.537420"}},{"identifier":{"@type":"URI","@value":"https://syndication.highwire.org/content/doi/10.1101/2023.04.20.537420"}}],"resourceType":"学術雑誌論文(journal article)","dc:title":[{"@value":"Efficient multiple gene knockout in\n                    <i>Colletotrichum higginsianum</i>\n                    via\n                    <scp>CRISPR/C</scp>\n                    as9 ribonucleoprotein and\n                    <i>URA3</i>\n                    ‐based marker recycling"}],"description":[{"type":"abstract","notation":[{"@value":"<jats:title>Abstract</jats:title>\n                  <jats:p>\n                    <jats:italic>Colletotrichum higginsianum</jats:italic>\n                    is a hemibiotrophic pathogen that causes anthracnose disease on crucifer hosts, including\n                    <jats:italic>Arabidopsis thaliana</jats:italic>\n                    . Despite the availability of genomic and transcriptomic information and the ability to transform both organisms, identifying\n                    <jats:italic>C. higginsianum</jats:italic>\n                    genes involved in virulence has been challenging due to recalcitrance to gene targeting and redundancy of virulence factors. To overcome these obstacles, we developed an efficient method for multiple gene disruption in\n                    <jats:italic>C. higginsianum</jats:italic>\n                    by combining CRISPR/Cas9 and a\n                    <jats:italic>URA3</jats:italic>\n                    ‐based marker recycling system. Our method significantly increased the efficiency of gene knockout via homologous recombination by introducing genomic DNA double‐strand breaks. We demonstrated the applicability of the\n                    <jats:italic>URA3</jats:italic>\n                    ‐based marker recycling system for multiple gene targeting in the same strain. Using our technology, we successfully targeted two melanin biosynthesis genes,\n                    <jats:italic>SCD1</jats:italic>\n                    and\n                    <jats:italic>PKS1</jats:italic>\n                    , which resulted in deficiency in melanization and loss of pathogenicity in the mutants. Our findings demonstrate the effectiveness of our methods in analysing virulence factors in\n                    <jats:italic>C. higginsianum</jats:italic>\n                    , thus accelerating research on plant–fungus interactions.\n                  </jats:p>"}]}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1380021391864693892","@type":"Researcher","foaf:name":[{"@value":"Katsuma Yonehara"}],"jpcoar:affiliationName":[{"@value":"RIKEN Center for Sustainable Resource Science  Yokohama Japan"},{"@value":"Department of Biological Science, Graduate School of Science The University of Tokyo  Tokyo Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1420282801210023680","@type":"Researcher","personIdentifier":[{"@type":"KAKEN_RESEARCHERS","@value":"50815438"},{"@type":"NRID","@value":"1000050815438"},{"@type":"NRID","@value":"9000408681749"},{"@type":"NRID","@value":"9000408613192"},{"@type":"NRID","@value":"9000350653323"},{"@type":"NRID","@value":"9000327926961"},{"@type":"RESEARCHMAP","@value":"https://researchmap.jp/kumakura827"}],"foaf:name":[{"@value":"Naoyoshi Kumakura"}],"jpcoar:affiliationName":[{"@value":"RIKEN Center for Sustainable Resource Science  Yokohama Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1420282801211444608","@type":"Researcher","personIdentifier":[{"@type":"KAKEN_RESEARCHERS","@value":"70291094"},{"@type":"NRID","@value":"1000070291094"},{"@type":"NRID","@value":"9000265559343"},{"@type":"NRID","@value":"9000398292461"},{"@type":"NRID","@value":"9000257963404"},{"@type":"NRID","@value":"9000398292483"}],"foaf:name":[{"@value":"Takayuki Motoyama"}],"jpcoar:affiliationName":[{"@value":"RIKEN Center for Sustainable Resource Science  Yokohama Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021391864693890","@type":"Researcher","foaf:name":[{"@value":"Nobuaki Ishihama"}],"jpcoar:affiliationName":[{"@value":"RIKEN Center for Sustainable Resource Science  Yokohama Japan"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021391864693893","@type":"Researcher","foaf:name":[{"@value":"Jean‐Félix Dallery"}],"jpcoar:affiliationName":[{"@value":"Université Paris‐Saclay, INRAE, UR BIOGER  Palaiseau France"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021391864693891","@type":"Researcher","foaf:name":[{"@value":"Richard O'Connell"}],"jpcoar:affiliationName":[{"@value":"Université Paris‐Saclay, INRAE, UR BIOGER  Palaiseau France"}]},{"@id":"https://cir.nii.ac.jp/crid/1380021391864693894","@type":"Researcher","foaf:name":[{"@value":"Ken Shirasu"}],"jpcoar:affiliationName":[{"@value":"RIKEN Center for Sustainable Resource Science  Yokohama Japan"},{"@value":"Department of Biological Science, Graduate School of Science The University of Tokyo  Tokyo Japan"}]}],"contributor":[{"@id":"https://cir.nii.ac.jp/crid/1891709542896121864","@type":"Researcher","foaf:name":[{"@value":"Agence japonaise pour la science et la technologie (JST) JPMJAX20B4"}]},{"@id":"https://cir.nii.ac.jp/crid/1891709542896121860","@type":"Researcher","foaf:name":[{"@value":"Ministère de l’éducation, de la culture, des sports, de la science et de la technologie, Japon (MEXT)Société japonaise pour la promotion de la science19KK0397"}]},{"@id":"https://cir.nii.ac.jp/crid/1891709542896121859","@type":"Researcher","foaf:name":[{"@value":"BIOlogie et GEstion des Risques en agriculture (BIOGER) ; Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)"}]},{"@id":"https://cir.nii.ac.jp/crid/1891709542896121856","@type":"Researcher","foaf:name":[{"@value":"RIKEN Center for Sustainable Resource Science [Yokohama] (RIKEN CSRS) ; RIKEN - Institute of Physical and Chemical Research [Japon] (RIKEN)"}]},{"@id":"https://cir.nii.ac.jp/crid/1891709542896121857","@type":"Researcher","foaf:name":[{"@value":"Department of Biological Sciences, Graduate School of Science, The University of Tokyo"}]}],"publication":{"publicationIdentifier":[{"@type":"PISSN","@value":"14646722"},{"@type":"EISSN","@value":"13643703"}],"prism:publicationName":[{"@value":"Molecular Plant Pathology"}],"dc:publisher":[{"@value":"Wiley"}],"prism:publicationDate":"2023-07-31","prism:volume":"24","prism:number":"11","prism:startingPage":"1451","prism:endingPage":"1464"},"reviewed":"false","dcterms:accessRights":"http://purl.org/coar/access_right/c_abf2","dc:rights":["http://creativecommons.org/licenses/by-nc-nd/4.0/","http://creativecommons.org/licenses/by-nc-nd/4.0/"],"url":[{"@id":"https://bsppjournals.onlinelibrary.wiley.com/doi/pdf/10.1111/mpp.13378"},{"@id":"https://syndication.highwire.org/content/doi/10.1101/2023.04.20.537420"}],"createdAt":"2023-07-31","modifiedAt":"2025-11-04","foaf:topic":[{"@id":"https://cir.nii.ac.jp/all?q=URA3","dc:title":"URA3"},{"@id":"https://cir.nii.ac.jp/all?q=Colletotrichum%20higginsianum","dc:title":"Colletotrichum higginsianum"},{"@id":"https://cir.nii.ac.jp/all?q=plant-pathogenic%20fungus","dc:title":"plant-pathogenic fungus"},{"@id":"https://cir.nii.ac.jp/all?q=Arabidopsis%20thaliana","dc:title":"Arabidopsis thaliana"},{"@id":"https://cir.nii.ac.jp/all?q=Technical%20Advance","dc:title":"Technical Advance"},{"@id":"https://cir.nii.ac.jp/all?q=%5BSDV%5DLife%20Sciences%20%5Bq-bio%5D","dc:title":"[SDV]Life Sciences [q-bio]"},{"@id":"https://cir.nii.ac.jp/all?q=%5BSDE%5DEnvironmental%20Sciences","dc:title":"[SDE]Environmental Sciences"},{"@id":"https://cir.nii.ac.jp/all?q=CRISPR/Cas9","dc:title":"CRISPR/Cas9"},{"@id":"https://cir.nii.ac.jp/all?q=selection%20marker%20recycling","dc:title":"selection marker recycling"}],"project":[{"@id":"https://cir.nii.ac.jp/crid/1040285300700225408","@type":"Project","projectIdentifier":[{"@type":"KAKEN","@value":"20K05820"},{"@type":"JGN","@value":"JP20K05820"},{"@type":"URI","@value":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20K05820/"}],"notation":[{"@language":"ja","@value":"「共生糸状菌」からの二次代謝産物発掘および生物間相互作用における役割の解明"},{"@language":"en","@value":"Mining of secondary metabolite from \"symbiotic fungi\" and elucidation of their roles in biological 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