Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering

Non-coding RNAs (crRNAs) produced from clustered regularly interspaced short palindromic repeats (CRISPR) loci and CRISPR-associated (Cas) proteins of the prokaryotic CRISPR-Cas systems form complexes that interfere with the spread of transmissible genetic elements through Cas-catalysed cleavage of...

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Detalles Bibliográficos
Autores: Esquerra-Ruvira, Belen, Baquedano, Ignacio, Ruiz, Raul, Fernández, Almudena, Montoliu, Lluís, Mojica, Francisco J M
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/363394
Acceso en línea:http://hdl.handle.net/10261/363394
https://api.elsevier.com/content/abstract/scopus_id/85153583624
Access Level:acceso abierto
Palabra clave:CRISPR
Cas9
genome editing
metagenome
recombineering
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spelling Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineeringEsquerra-Ruvira, BelenBaquedano, IgnacioRuiz, RaulFernández, AlmudenaMontoliu, LluísMojica, Francisco J MCRISPRCas9genome editingmetagenomerecombineeringNon-coding RNAs (crRNAs) produced from clustered regularly interspaced short palindromic repeats (CRISPR) loci and CRISPR-associated (Cas) proteins of the prokaryotic CRISPR-Cas systems form complexes that interfere with the spread of transmissible genetic elements through Cas-catalysed cleavage of foreign genetic material matching the guide crRNA sequences. The easily programmable targeting of nucleic acids enabled by these ribonucleoproteins has facilitated the implementation of CRISPR-based molecular biology tools for in vivo and in vitro modification of DNA and RNA targets. Despite the diversity of DNA-targeting Cas nucleases so far identified, native and engineered derivatives of the Streptococcus pyogenes SpCas9 are the most widely used for genome engineering, at least in part due to their catalytic robustness and the requirement of an exceptionally short motif (5'-NGG-3' PAM) flanking the target sequence. However, the large size of the SpCas9 variants impairs the delivery of the tool to eukaryotic cells and smaller alternatives are desirable. Here, we identify in a metagenome a new CRISPR-Cas9 system associated with a smaller Cas9 protein (EHCas9) that targets DNA sequences flanked by 5'-NGG-3' PAMs. We develop a simplified EHCas9 tool that specifically cleaves DNA targets and is functional for genome editing applications in prokaryotes and eukaryotic cells.Conselleria de Innovación, Universidades, Ciencia y Sociedad Digital, Generalitat Valenciana (CIUCSD), Grant/Award Numbers: PROMETEO/2017/129 and PROMETEO/2021/057. Ministerio de Ciencia e Innovación (MICINN), Grant/Award Numbers: RTI2018- 101223-B-I00 and PID2021-127644OB-I00.Peer reviewedJohn Wiley & SonsGeneralitat ValencianaMinisterio de Ciencia e Innovación (España)Montoliu, Lluís [0000-0003-3941-1176]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/363394https://api.elsevier.com/content/abstract/scopus_id/85153583624reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-101223-B-I00info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127644OB-I00Microbial biotechnologyapplication/pdfhttps://enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.14266Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3633942026-05-22T06:33:51Z
dc.title.none.fl_str_mv Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
title Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
spellingShingle Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
Esquerra-Ruvira, Belen
CRISPR
Cas9
genome editing
metagenome
recombineering
title_short Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
title_full Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
title_fullStr Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
title_full_unstemmed Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
title_sort Identification of the EH CRISPR-Cas9 system on a metagenome and its application to genome engineering
dc.creator.none.fl_str_mv Esquerra-Ruvira, Belen
Baquedano, Ignacio
Ruiz, Raul
Fernández, Almudena
Montoliu, Lluís
Mojica, Francisco J M
author Esquerra-Ruvira, Belen
author_facet Esquerra-Ruvira, Belen
Baquedano, Ignacio
Ruiz, Raul
Fernández, Almudena
Montoliu, Lluís
Mojica, Francisco J M
author_role author
author2 Baquedano, Ignacio
Ruiz, Raul
Fernández, Almudena
Montoliu, Lluís
Mojica, Francisco J M
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Generalitat Valenciana
Ministerio de Ciencia e Innovación (España)
Montoliu, Lluís [0000-0003-3941-1176]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv CRISPR
Cas9
genome editing
metagenome
recombineering
topic CRISPR
Cas9
genome editing
metagenome
recombineering
description Non-coding RNAs (crRNAs) produced from clustered regularly interspaced short palindromic repeats (CRISPR) loci and CRISPR-associated (Cas) proteins of the prokaryotic CRISPR-Cas systems form complexes that interfere with the spread of transmissible genetic elements through Cas-catalysed cleavage of foreign genetic material matching the guide crRNA sequences. The easily programmable targeting of nucleic acids enabled by these ribonucleoproteins has facilitated the implementation of CRISPR-based molecular biology tools for in vivo and in vitro modification of DNA and RNA targets. Despite the diversity of DNA-targeting Cas nucleases so far identified, native and engineered derivatives of the Streptococcus pyogenes SpCas9 are the most widely used for genome engineering, at least in part due to their catalytic robustness and the requirement of an exceptionally short motif (5'-NGG-3' PAM) flanking the target sequence. However, the large size of the SpCas9 variants impairs the delivery of the tool to eukaryotic cells and smaller alternatives are desirable. Here, we identify in a metagenome a new CRISPR-Cas9 system associated with a smaller Cas9 protein (EHCas9) that targets DNA sequences flanked by 5'-NGG-3' PAMs. We develop a simplified EHCas9 tool that specifically cleaves DNA targets and is functional for genome editing applications in prokaryotes and eukaryotic cells.
publishDate 2023
dc.date.none.fl_str_mv 2023
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/363394
https://api.elsevier.com/content/abstract/scopus_id/85153583624
url http://hdl.handle.net/10261/363394
https://api.elsevier.com/content/abstract/scopus_id/85153583624
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-101223-B-I00
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127644OB-I00
Microbial biotechnology
application/pdf
https://enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.14266
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv John Wiley & Sons
publisher.none.fl_str_mv John Wiley & Sons
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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