CRISPR-on system for the activation of the endogenous human INS gene

Autores
Giménez, Carla Alejandra; Ielpi, Marcelo; Mutto, Adrián Angel; Grosembacher, Luis; Argibay, Pablo; Pereyra Bonnet, Federico Alberto
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Advances in the field of epigenetics have allowed the design of new therapeutic strategies to address complex diseases such as type 1 diabetes (T1D). Clustered regularly interspaced short palindromic repeats (CRISPR)-on is a novel and powerful RNA-guided transcriptional activator system that can turn on specific gene expression; however, it remains unclear whether this system can be widely used or whether its use will be restricted depending on cell types, methylation promoter statuses or the capacity to modulate chromatin state. Our results revealed that the CRISPR-on system fused with transcriptional activators (dCas9-VP160) activated endogenous human INS, which is a silenced gene with a fully methylated promoter. Similarly, we observed a synergistic effect on gene activation when multiple single guide RNAs were used, and the transcriptional activation was maintained until day 21. Regarding the epigenetic profile, the targeted promoter gene did not exhibit alteration in its methylation status but rather exhibited altered levels of H3K9ac following treatment. Importantly, we showed that dCas9-VP160 acts on patients' cells in vitro, particularly the fibroblasts of patients with T1D.
Fil: Giménez, Carla Alejandra. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ielpi, Marcelo. Hospital Italiano; Argentina
Fil: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Grosembacher, Luis. Hospital Italiano; Argentina
Fil: Argibay, Pablo. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Pereyra Bonnet, Federico Alberto. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
Crisprs
Rna
Esc
Insulin
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/44026
Acceder
id CONICETDig_e5fee60703ab4bd14c420aaee546ceb1
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling CRISPR-on system for the activation of the endogenous human INS geneGiménez, Carla AlejandraIelpi, MarceloMutto, Adrián AngelGrosembacher, LuisArgibay, PabloPereyra Bonnet, Federico AlbertoCrisprsRnaEscInsulinhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Advances in the field of epigenetics have allowed the design of new therapeutic strategies to address complex diseases such as type 1 diabetes (T1D). Clustered regularly interspaced short palindromic repeats (CRISPR)-on is a novel and powerful RNA-guided transcriptional activator system that can turn on specific gene expression; however, it remains unclear whether this system can be widely used or whether its use will be restricted depending on cell types, methylation promoter statuses or the capacity to modulate chromatin state. Our results revealed that the CRISPR-on system fused with transcriptional activators (dCas9-VP160) activated endogenous human INS, which is a silenced gene with a fully methylated promoter. Similarly, we observed a synergistic effect on gene activation when multiple single guide RNAs were used, and the transcriptional activation was maintained until day 21. Regarding the epigenetic profile, the targeted promoter gene did not exhibit alteration in its methylation status but rather exhibited altered levels of H3K9ac following treatment. Importantly, we showed that dCas9-VP160 acts on patients' cells in vitro, particularly the fibroblasts of patients with T1D.Fil: Giménez, Carla Alejandra. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ielpi, Marcelo. Hospital Italiano; ArgentinaFil: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Grosembacher, Luis. Hospital Italiano; ArgentinaFil: Argibay, Pablo. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pereyra Bonnet, Federico Alberto. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaNature Publishing Group2016-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/44026Giménez, Carla Alejandra; Ielpi, Marcelo; Mutto, Adrián Angel; Grosembacher, Luis; Argibay, Pablo; et al.; CRISPR-on system for the activation of the endogenous human INS gene; Nature Publishing Group; Gene Therapy; 23; 4-2016; 543–5470969-7128CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1038/gt.2016.28info:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/gt201628info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:41:38Zoai:ri.conicet.gov.ar:11336/44026instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:41:38.772CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv CRISPR-on system for the activation of the endogenous human INS gene
title CRISPR-on system for the activation of the endogenous human INS gene
spellingShingle CRISPR-on system for the activation of the endogenous human INS gene
Giménez, Carla Alejandra
Crisprs
Rna
Esc
Insulin
title_short CRISPR-on system for the activation of the endogenous human INS gene
title_full CRISPR-on system for the activation of the endogenous human INS gene
title_fullStr CRISPR-on system for the activation of the endogenous human INS gene
title_full_unstemmed CRISPR-on system for the activation of the endogenous human INS gene
title_sort CRISPR-on system for the activation of the endogenous human INS gene
dc.creator.none.fl_str_mv Giménez, Carla Alejandra
Ielpi, Marcelo
Mutto, Adrián Angel
Grosembacher, Luis
Argibay, Pablo
Pereyra Bonnet, Federico Alberto
author Giménez, Carla Alejandra
author_facet Giménez, Carla Alejandra
Ielpi, Marcelo
Mutto, Adrián Angel
Grosembacher, Luis
Argibay, Pablo
Pereyra Bonnet, Federico Alberto
author_role author
author2 Ielpi, Marcelo
Mutto, Adrián Angel
Grosembacher, Luis
Argibay, Pablo
Pereyra Bonnet, Federico Alberto
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Crisprs
Rna
Esc
Insulin
topic Crisprs
Rna
Esc
Insulin
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Advances in the field of epigenetics have allowed the design of new therapeutic strategies to address complex diseases such as type 1 diabetes (T1D). Clustered regularly interspaced short palindromic repeats (CRISPR)-on is a novel and powerful RNA-guided transcriptional activator system that can turn on specific gene expression; however, it remains unclear whether this system can be widely used or whether its use will be restricted depending on cell types, methylation promoter statuses or the capacity to modulate chromatin state. Our results revealed that the CRISPR-on system fused with transcriptional activators (dCas9-VP160) activated endogenous human INS, which is a silenced gene with a fully methylated promoter. Similarly, we observed a synergistic effect on gene activation when multiple single guide RNAs were used, and the transcriptional activation was maintained until day 21. Regarding the epigenetic profile, the targeted promoter gene did not exhibit alteration in its methylation status but rather exhibited altered levels of H3K9ac following treatment. Importantly, we showed that dCas9-VP160 acts on patients' cells in vitro, particularly the fibroblasts of patients with T1D.
Fil: Giménez, Carla Alejandra. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ielpi, Marcelo. Hospital Italiano; Argentina
Fil: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Grosembacher, Luis. Hospital Italiano; Argentina
Fil: Argibay, Pablo. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Pereyra Bonnet, Federico Alberto. Hospital Italiano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Advances in the field of epigenetics have allowed the design of new therapeutic strategies to address complex diseases such as type 1 diabetes (T1D). Clustered regularly interspaced short palindromic repeats (CRISPR)-on is a novel and powerful RNA-guided transcriptional activator system that can turn on specific gene expression; however, it remains unclear whether this system can be widely used or whether its use will be restricted depending on cell types, methylation promoter statuses or the capacity to modulate chromatin state. Our results revealed that the CRISPR-on system fused with transcriptional activators (dCas9-VP160) activated endogenous human INS, which is a silenced gene with a fully methylated promoter. Similarly, we observed a synergistic effect on gene activation when multiple single guide RNAs were used, and the transcriptional activation was maintained until day 21. Regarding the epigenetic profile, the targeted promoter gene did not exhibit alteration in its methylation status but rather exhibited altered levels of H3K9ac following treatment. Importantly, we showed that dCas9-VP160 acts on patients' cells in vitro, particularly the fibroblasts of patients with T1D.
publishDate 2016
dc.date.none.fl_str_mv 2016-04
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/44026
Giménez, Carla Alejandra; Ielpi, Marcelo; Mutto, Adrián Angel; Grosembacher, Luis; Argibay, Pablo; et al.; CRISPR-on system for the activation of the endogenous human INS gene; Nature Publishing Group; Gene Therapy; 23; 4-2016; 543–547
0969-7128
CONICET Digital
CONICET
url http://hdl.handle.net/11336/44026
identifier_str_mv Giménez, Carla Alejandra; Ielpi, Marcelo; Mutto, Adrián Angel; Grosembacher, Luis; Argibay, Pablo; et al.; CRISPR-on system for the activation of the endogenous human INS gene; Nature Publishing Group; Gene Therapy; 23; 4-2016; 543–547
0969-7128
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1038/gt.2016.28
info:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/gt201628
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844613313819836416
score 13.070432

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