A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System

The RNA-guided endonuclease Cas9 (CRISPR-Cas9) genome editing system has been widely used for biomedical research and holds great potential for therapeutic applications in eukaryotes. The conventional vector-based CRISPR-Cas9 delivery system requires two different RNA polymerase promoters for expres...

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Detalles Bibliográficos
Autores: Gao, Zongliang, Herrera, Elena, Berkhout, Ben
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/415230
Acceso en línea:http://hdl.handle.net/10261/415230
https://api.elsevier.com/content/abstract/scopus_id/85057736079
Access Level:acceso abierto
Palabra clave:CRISPR-Cas9
Cas9 protein expression
H1 promoter
RNA polymerase III and II
Dual RNA polymerase promoter
Guide RNA expression
Lentiviral vector
Viral delivery
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spelling A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 SystemGao, ZongliangHerrera, ElenaBerkhout, BenCRISPR-Cas9Cas9 protein expressionH1 promoterRNA polymerase III and IIDual RNA polymerase promoterGuide RNA expressionLentiviral vectorViral deliveryThe RNA-guided endonuclease Cas9 (CRISPR-Cas9) genome editing system has been widely used for biomedical research and holds great potential for therapeutic applications in eukaryotes. The conventional vector-based CRISPR-Cas9 delivery system requires two different RNA polymerase promoters for expression of the guide RNA (gRNA) and Cas9 endonuclease. The large size and relative complexity of such CRISPR transgene cassettes impede their broad implementation, especially in gene therapy applications with viral vectors that have a limited packaging capacity. Here, we report the design of a single-promoter-driven CRISPR-Cas9 system that uses the dual-polymerase (Pol II and Pol III) activity of the H1 promoter. This size reduction strategy of the vector insert provides a significant titer advantage in the lentiviral vector over the regular CRISPR system.This research was supported by the AIDS Fonds (HIV– P-37102). Z.G. is supported by a scholarship from the China Scholarship Council (CSC). We thank Yi Zheng for the kind donation of reagents.Peer reviewedElsevierAids FondsChina Scholarship CouncilHerrera Carrillo, Elena [0000-0001-9986-8552]202620262019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/415230https://api.elsevier.com/content/abstract/scopus_id/85057736079reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésThe underlying dataset has been published as supplementary material of the article in the publisher platform at DOI 10.1016/j.omtn.2018.10.016https://doi.org/10.1016/j.omtn.2018.10.016Noinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4152302026-05-22T06:33:51Z
dc.title.none.fl_str_mv A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
title A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
spellingShingle A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
Gao, Zongliang
CRISPR-Cas9
Cas9 protein expression
H1 promoter
RNA polymerase III and II
Dual RNA polymerase promoter
Guide RNA expression
Lentiviral vector
Viral delivery
title_short A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
title_full A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
title_fullStr A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
title_full_unstemmed A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
title_sort A Single H1 Promoter Can Drive Both Guide RNA and Endonuclease Expression in the CRISPR-Cas9 System
dc.creator.none.fl_str_mv Gao, Zongliang
Herrera, Elena
Berkhout, Ben
author Gao, Zongliang
author_facet Gao, Zongliang
Herrera, Elena
Berkhout, Ben
author_role author
author2 Herrera, Elena
Berkhout, Ben
author2_role author
author
dc.contributor.none.fl_str_mv Aids Fonds
China Scholarship Council
Herrera Carrillo, Elena [0000-0001-9986-8552]
dc.subject.none.fl_str_mv CRISPR-Cas9
Cas9 protein expression
H1 promoter
RNA polymerase III and II
Dual RNA polymerase promoter
Guide RNA expression
Lentiviral vector
Viral delivery
topic CRISPR-Cas9
Cas9 protein expression
H1 promoter
RNA polymerase III and II
Dual RNA polymerase promoter
Guide RNA expression
Lentiviral vector
Viral delivery
description The RNA-guided endonuclease Cas9 (CRISPR-Cas9) genome editing system has been widely used for biomedical research and holds great potential for therapeutic applications in eukaryotes. The conventional vector-based CRISPR-Cas9 delivery system requires two different RNA polymerase promoters for expression of the guide RNA (gRNA) and Cas9 endonuclease. The large size and relative complexity of such CRISPR transgene cassettes impede their broad implementation, especially in gene therapy applications with viral vectors that have a limited packaging capacity. Here, we report the design of a single-promoter-driven CRISPR-Cas9 system that uses the dual-polymerase (Pol II and Pol III) activity of the H1 promoter. This size reduction strategy of the vector insert provides a significant titer advantage in the lentiviral vector over the regular CRISPR system.
publishDate 2019
dc.date.none.fl_str_mv 2019
2026
2026
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/10261/415230
https://api.elsevier.com/content/abstract/scopus_id/85057736079
url http://hdl.handle.net/10261/415230
https://api.elsevier.com/content/abstract/scopus_id/85057736079
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI 10.1016/j.omtn.2018.10.016
https://doi.org/10.1016/j.omtn.2018.10.016
No
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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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