Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection

Allogeneic transplantation of blood stem cells from a CCR5-Δ32 homozygous donor to an HIV-infected individual, the "Berlin patient", led to a cure. Since then there has been a search for approaches that mimic this intervention in a gene therapy setting. RNA interference (RNAi) has evolved...

ver descrição completa

Detalhes bibliográficos
Autores: Herrera, Elena, Berkhout, Ben
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2017
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/415342
Acesso em linha:http://hdl.handle.net/10261/415342
https://api.elsevier.com/content/abstract/scopus_id/85019679237
Access Level:acceso abierto
Palavra-chave:RNAi
Dicer
Ago2
shRNA design
Dicer-independent shRNA
miR-451
CCR5
id ES_791d7f2b6d8df2c8149d0c5063293b31
oai_identifier_str oai:digital.csic.es:10261/415342
network_acronym_str ES
network_name_str España
repository_id_str
spelling Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infectionHerrera, ElenaBerkhout, BenRNAiDicerAgo2shRNA designDicer-independent shRNAmiR-451CCR5Allogeneic transplantation of blood stem cells from a CCR5-Δ32 homozygous donor to an HIV-infected individual, the "Berlin patient", led to a cure. Since then there has been a search for approaches that mimic this intervention in a gene therapy setting. RNA interference (RNAi) has evolved as a powerful tool to regulate gene expression in a sequence-specific manner and can be used to inactivate the CCR5 mRNA. Short hairpin RNA (shRNA) molecules can impair CCR5 expression, but these molecules may cause unintended side effects and they will not be processed in cells that lack Dicer, such as monocytes. Dicer-independent RNAi pathways have opened opportunities for new AgoshRNA designs that rely exclusively on Ago2 for maturation. Furthermore, AgoshRNA processing yields a single active guide RNA, thus reducing off-target effects. In this study, we tested different AgoshRNA designs against CCR5. We selected AgoshRNAs that potently downregulated CCR5 expression on human T cells and peripheral blood mononuclear cells (PBMC) and that had no apparent adverse effect on T cell development as assessed in a competitive cell growth assay. CCR5 knockdown significantly protected T cells from CCR5 tropic HIV-1 infection.This work was supported by the Nederlandse Organisatie voor Wetenschappelijk Onderzoek—Chemische Wetenschappen (NWO-CW, Top Grant) and Zorg Onderzoek Nederland—Medische Wetenschappen (ZonMw, Translational Gene Therapy Grant). We thank Berend Hooibrink for expertise in cell sorting and maintenance of the flow cytometry facility. The authors declare no conflict of interest.Peer reviewedPublic Library of ScienceDutch Research CouncilZonMw202620262017info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/415342https://api.elsevier.com/content/abstract/scopus_id/85019679237reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.1371/journal.pone.0177935Noinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4153422026-05-22T06:33:51Z
dc.title.none.fl_str_mv Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
title Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
spellingShingle Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
Herrera, Elena
RNAi
Dicer
Ago2
shRNA design
Dicer-independent shRNA
miR-451
CCR5
title_short Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
title_full Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
title_fullStr Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
title_full_unstemmed Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
title_sort Novel AgoshRNA molecules for silencing of the CCR5 co-receptor for HIV-1 infection
dc.creator.none.fl_str_mv Herrera, Elena
Berkhout, Ben
author Herrera, Elena
author_facet Herrera, Elena
Berkhout, Ben
author_role author
author2 Berkhout, Ben
author2_role author
dc.contributor.none.fl_str_mv Dutch Research Council
ZonMw
dc.subject.none.fl_str_mv RNAi
Dicer
Ago2
shRNA design
Dicer-independent shRNA
miR-451
CCR5
topic RNAi
Dicer
Ago2
shRNA design
Dicer-independent shRNA
miR-451
CCR5
description Allogeneic transplantation of blood stem cells from a CCR5-Δ32 homozygous donor to an HIV-infected individual, the "Berlin patient", led to a cure. Since then there has been a search for approaches that mimic this intervention in a gene therapy setting. RNA interference (RNAi) has evolved as a powerful tool to regulate gene expression in a sequence-specific manner and can be used to inactivate the CCR5 mRNA. Short hairpin RNA (shRNA) molecules can impair CCR5 expression, but these molecules may cause unintended side effects and they will not be processed in cells that lack Dicer, such as monocytes. Dicer-independent RNAi pathways have opened opportunities for new AgoshRNA designs that rely exclusively on Ago2 for maturation. Furthermore, AgoshRNA processing yields a single active guide RNA, thus reducing off-target effects. In this study, we tested different AgoshRNA designs against CCR5. We selected AgoshRNAs that potently downregulated CCR5 expression on human T cells and peripheral blood mononuclear cells (PBMC) and that had no apparent adverse effect on T cell development as assessed in a competitive cell growth assay. CCR5 knockdown significantly protected T cells from CCR5 tropic HIV-1 infection.
publishDate 2017
dc.date.none.fl_str_mv 2017
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/10261/415342
https://api.elsevier.com/content/abstract/scopus_id/85019679237
url http://hdl.handle.net/10261/415342
https://api.elsevier.com/content/abstract/scopus_id/85019679237
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.1371/journal.pone.0177935
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 Public Library of Science
publisher.none.fl_str_mv Public Library of Science
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
_version_ 1869411323816706048
score 15,228081

Registos relacionados