CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells

Conventional reprogramming methods rely on the ectopic expression of transcription factors to reprogram somatic cells into induced pluripotent stem cells (iPSCs). The forced expression of transcription factors may lead to off-target gene activation and heterogeneous reprogramming, resulting in the e...

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Autores: Sokka, Joonas, Yoshihara, Masahito, Kvist, Jouni, Laiho, Laura, Warren, Andrew, Stadelmann, Christian, Jouhilahti, Eeva-Mari, Kilpinen, Helena, Balboa, Diego, Katayama, Shintaro, Kyttälä, Aija, Kere, Juha, Otonkoski, Timo, Weltner, Jere, Trokovic, Ras
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Universitat Pompeu Fabra
Repositorio:Repositorio Digital de la UPF
OAI Identifier:oai:repositori.upf.edu:10230/52856
Acceso en línea:http://hdl.handle.net/10230/52856
http://dx.doi.org/10.1016/j.stemcr.2021.12.017
Access Level:acceso abierto
Palabra clave:CRISPRa
EEA
LCL
Human
iPSC
microRNA
mir-302/367
Reprogramming
Single-cell RNA sequencing
Transcriptomics
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spelling CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cellsSokka, JoonasYoshihara, MasahitoKvist, JouniLaiho, LauraWarren, AndrewStadelmann, ChristianJouhilahti, Eeva-MariKilpinen, HelenaBalboa, DiegoKatayama, ShintaroKyttälä, AijaKere, JuhaOtonkoski, TimoWeltner, JereTrokovic, RasCRISPRaEEALCLHumaniPSCmicroRNAmir-302/367ReprogrammingSingle-cell RNA sequencingTranscriptomicsConventional reprogramming methods rely on the ectopic expression of transcription factors to reprogram somatic cells into induced pluripotent stem cells (iPSCs). The forced expression of transcription factors may lead to off-target gene activation and heterogeneous reprogramming, resulting in the emergence of alternative cell types and aberrant iPSCs. Activation of endogenous pluripotency factors by CRISPR activation (CRISPRa) can reduce this heterogeneity. Here, we describe a high-efficiency reprogramming of human somatic cells into iPSCs using optimized CRISPRa. Efficient reprogramming was dependent on the additional targeting of the embryo genome activation-enriched Alu-motif and the miR-302/367 locus. Single-cell transcriptome analysis revealed that the optimized CRISPRa reprogrammed cells more directly and specifically into the pluripotent state when compared to the conventional reprogramming method. These findings support the use of CRISPRa for high-quality pluripotent reprogramming of human cells.Elsevier202220222022info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/52856http://dx.doi.org/10.1016/j.stemcr.2021.12.017reponame:Repositorio Digital de la UPFinstname:Universitat Pompeu FabraInglésStem Cell Reports. 2022 Feb 8;17(2):413-26© 2021 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositori.upf.edu:10230/528562026-06-12T07:21:37Z
dc.title.none.fl_str_mv CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
title CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
spellingShingle CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
Sokka, Joonas
CRISPRa
EEA
LCL
Human
iPSC
microRNA
mir-302/367
Reprogramming
Single-cell RNA sequencing
Transcriptomics
title_short CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
title_full CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
title_fullStr CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
title_full_unstemmed CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
title_sort CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells
dc.creator.none.fl_str_mv Sokka, Joonas
Yoshihara, Masahito
Kvist, Jouni
Laiho, Laura
Warren, Andrew
Stadelmann, Christian
Jouhilahti, Eeva-Mari
Kilpinen, Helena
Balboa, Diego
Katayama, Shintaro
Kyttälä, Aija
Kere, Juha
Otonkoski, Timo
Weltner, Jere
Trokovic, Ras
author Sokka, Joonas
author_facet Sokka, Joonas
Yoshihara, Masahito
Kvist, Jouni
Laiho, Laura
Warren, Andrew
Stadelmann, Christian
Jouhilahti, Eeva-Mari
Kilpinen, Helena
Balboa, Diego
Katayama, Shintaro
Kyttälä, Aija
Kere, Juha
Otonkoski, Timo
Weltner, Jere
Trokovic, Ras
author_role author
author2 Yoshihara, Masahito
Kvist, Jouni
Laiho, Laura
Warren, Andrew
Stadelmann, Christian
Jouhilahti, Eeva-Mari
Kilpinen, Helena
Balboa, Diego
Katayama, Shintaro
Kyttälä, Aija
Kere, Juha
Otonkoski, Timo
Weltner, Jere
Trokovic, Ras
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CRISPRa
EEA
LCL
Human
iPSC
microRNA
mir-302/367
Reprogramming
Single-cell RNA sequencing
Transcriptomics
topic CRISPRa
EEA
LCL
Human
iPSC
microRNA
mir-302/367
Reprogramming
Single-cell RNA sequencing
Transcriptomics
description Conventional reprogramming methods rely on the ectopic expression of transcription factors to reprogram somatic cells into induced pluripotent stem cells (iPSCs). The forced expression of transcription factors may lead to off-target gene activation and heterogeneous reprogramming, resulting in the emergence of alternative cell types and aberrant iPSCs. Activation of endogenous pluripotency factors by CRISPR activation (CRISPRa) can reduce this heterogeneity. Here, we describe a high-efficiency reprogramming of human somatic cells into iPSCs using optimized CRISPRa. Efficient reprogramming was dependent on the additional targeting of the embryo genome activation-enriched Alu-motif and the miR-302/367 locus. Single-cell transcriptome analysis revealed that the optimized CRISPRa reprogrammed cells more directly and specifically into the pluripotent state when compared to the conventional reprogramming method. These findings support the use of CRISPRa for high-quality pluripotent reprogramming of human cells.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/52856
http://dx.doi.org/10.1016/j.stemcr.2021.12.017
url http://hdl.handle.net/10230/52856
http://dx.doi.org/10.1016/j.stemcr.2021.12.017
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Stem Cell Reports. 2022 Feb 8;17(2):413-26
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositorio Digital de la UPF
instname:Universitat Pompeu Fabra
instname_str Universitat Pompeu Fabra
reponame_str Repositorio Digital de la UPF
collection Repositorio Digital de la UPF
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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