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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Detalles Bibliográficos
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:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat: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
Descripción
Sumario: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.

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