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...
| Autores: | ; ; ; ; ; ; ; ; ; ; ; ; ; ; |
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| Tipo de documento: | Artículo |
| Estado: | Published version |
| 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 |
| Nivel de acceso: | Acceso abierto |
| Palabra clave: | CRISPRa EEA LCL Human iPSC microRNA mir-302/367 Reprogramming Single-cell RNA sequencing Transcriptomics |
| _version_ | 1878729522254184448 |
|---|---|
| author | Sokka, Joonas |
| 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 |
| author_browse | Balboa, Diego Jouhilahti, Eeva-Mari Katayama, Shintaro Kere, Juha Kilpinen, Helena Kvist, Jouni Kyttälä, Aija Laiho, Laura Otonkoski, Timo Sokka, Joonas Stadelmann, Christian Trokovic, Ras Warren, Andrew Weltner, Jere Yoshihara, Masahito |
| 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 |
| collection | Repositorio Digital de la UPF |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_099eafc8953c722d9e0da351ffc19c98 |
| institution | Universitat Pompeu Fabra |
| instname_str | Universitat Pompeu Fabra |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:repositori.upf.edu:10230/52856 |
| publishDate | 2022 |
| publishDateSort | 2022 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| reponame_str | Repositorio Digital de la UPF |
| spelling | CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells 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 CRISPRa EEA LCL Human iPSC microRNA mir-302/367 Reprogramming Single-cell RNA sequencing Transcriptomics 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. Elsevier http://hdl.handle.net/10230/52856 http://dx.doi.org/10.1016/j.stemcr.2021.12.017 |
| 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 |
| status_str | publishedVersion |
| title | 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_short | CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells |
| title_sort | CRISPR activation enables high-fidelity reprogramming into human pluripotent stem cells |
| topic | CRISPRa EEA LCL Human iPSC microRNA mir-302/367 Reprogramming Single-cell RNA sequencing Transcriptomics |
| topic_facet | CRISPRa EEA LCL Human iPSC microRNA mir-302/367 Reprogramming Single-cell RNA sequencing Transcriptomics |
| url | http://hdl.handle.net/10230/52856 http://dx.doi.org/10.1016/j.stemcr.2021.12.017 |