Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity

RNA polymerase III (Pol III) promoters, such as 7SK, U6, and H1, are widely used for the expression of small noncoding RNAs, including short hairpin RNAs for RNAi experiments and guide RNAs for CRISPR-mediated genome editing. We previously reported dual RNA polymerase activity (Pol II/III) for the h...

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Detalles Bibliográficos
Autores: Gao, Zongliang, Velden, Yme van der, Fan, Minghui, Linden, Cynthia Alyssa van der, Vink, Monique, Herrera, Elena, Berkhout, Ben
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/405741
Acceso en línea:http://hdl.handle.net/10261/405741
https://api.elsevier.com/content/abstract/scopus_id/85102940590
Access Level:acceso abierto
Palabra clave:H1 promoter
Pol II
Pol III
RNA polymerase
Engineering
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spelling Engineered miniature H1 promoters with dedicated RNA polymerase II or III activityGao, ZongliangVelden, Yme van derFan, MinghuiLinden, Cynthia Alyssa van derVink, MoniqueHerrera, ElenaBerkhout, BenH1 promoterPol IIPol IIIRNA polymeraseEngineeringRNA polymerase III (Pol III) promoters, such as 7SK, U6, and H1, are widely used for the expression of small noncoding RNAs, including short hairpin RNAs for RNAi experiments and guide RNAs for CRISPR-mediated genome editing. We previously reported dual RNA polymerase activity (Pol II/III) for the human H1 promoter and demonstrated that this promiscuous RNA polymerase use can be exploited for the simultaneous expression of both a noncoding RNA and an mRNA. However, this combination is not a desired feature in other experimental and therapeutic settings. To overcome this limitation of the H1 promoter, we engineered a miniature H1/7SK hybrid promoter with minimal Pol II activity, thereby boosting Pol III activity to a level that is higher than that of either parental promoter. In parallel, we also engineered small Pol II-specific H1 promoter variants and explored their use as general Pol II promoters for protein expression. The newly engineered promoter variants form an attractive alternative to the commonly used H1 promoter in terms of not only activity and small promoter size but also concerning safety by exclusive expression of the desired therapeutic transcript (either pol II or pol III but not both).This research was supported by National Institutes of Health (RO1 grant: 1R01AI145045IH to B. B., E. H.-C.), AIDS Fonds grant (HIV-P-37102 to E. H.-C.) and King Baudouin Foundation (to B. B.). Z. G. and M. F. are supported by a scholarship from the China Scholarship Council.NoAmerican Society for Biochemistry and Molecular BiologyElsevierNational Institutes of Health (US)Aids FondsKing Baudouin Foundation United StatesChina Scholarship CouncilHerrera, Elena [0000-0001-9986-8552]202520252021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/405741https://api.elsevier.com/content/abstract/scopus_id/85102940590reponame: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 10.1074/jbc.RA120.015386https://doi.org/10.1074/jbc.RA120.015386Noinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4057412026-05-22T06:33:51Z
dc.title.none.fl_str_mv Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
title Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
spellingShingle Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
Gao, Zongliang
H1 promoter
Pol II
Pol III
RNA polymerase
Engineering
title_short Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
title_full Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
title_fullStr Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
title_full_unstemmed Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
title_sort Engineered miniature H1 promoters with dedicated RNA polymerase II or III activity
dc.creator.none.fl_str_mv Gao, Zongliang
Velden, Yme van der
Fan, Minghui
Linden, Cynthia Alyssa van der
Vink, Monique
Herrera, Elena
Berkhout, Ben
author Gao, Zongliang
author_facet Gao, Zongliang
Velden, Yme van der
Fan, Minghui
Linden, Cynthia Alyssa van der
Vink, Monique
Herrera, Elena
Berkhout, Ben
author_role author
author2 Velden, Yme van der
Fan, Minghui
Linden, Cynthia Alyssa van der
Vink, Monique
Herrera, Elena
Berkhout, Ben
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv National Institutes of Health (US)
Aids Fonds
King Baudouin Foundation United States
China Scholarship Council
Herrera, Elena [0000-0001-9986-8552]
dc.subject.none.fl_str_mv H1 promoter
Pol II
Pol III
RNA polymerase
Engineering
topic H1 promoter
Pol II
Pol III
RNA polymerase
Engineering
description RNA polymerase III (Pol III) promoters, such as 7SK, U6, and H1, are widely used for the expression of small noncoding RNAs, including short hairpin RNAs for RNAi experiments and guide RNAs for CRISPR-mediated genome editing. We previously reported dual RNA polymerase activity (Pol II/III) for the human H1 promoter and demonstrated that this promiscuous RNA polymerase use can be exploited for the simultaneous expression of both a noncoding RNA and an mRNA. However, this combination is not a desired feature in other experimental and therapeutic settings. To overcome this limitation of the H1 promoter, we engineered a miniature H1/7SK hybrid promoter with minimal Pol II activity, thereby boosting Pol III activity to a level that is higher than that of either parental promoter. In parallel, we also engineered small Pol II-specific H1 promoter variants and explored their use as general Pol II promoters for protein expression. The newly engineered promoter variants form an attractive alternative to the commonly used H1 promoter in terms of not only activity and small promoter size but also concerning safety by exclusive expression of the desired therapeutic transcript (either pol II or pol III but not both).
publishDate 2021
dc.date.none.fl_str_mv 2021
2025
2025
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/10261/405741
https://api.elsevier.com/content/abstract/scopus_id/85102940590
url http://hdl.handle.net/10261/405741
https://api.elsevier.com/content/abstract/scopus_id/85102940590
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 10.1074/jbc.RA120.015386
https://doi.org/10.1074/jbc.RA120.015386
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 American Society for Biochemistry and Molecular Biology
Elsevier
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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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