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Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x.

Human pluripotent reprogramming with CRISPR activators.

Author information

1
Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, Helsinki, 00014, Finland. jere.weltner@helsinki.fi.
2
Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, Helsinki, 00014, Finland.
3
Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, 141 83, Sweden.
4
Competence Centre on Health Technologies, Tartu, 50410, Estonia.
5
Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, Helsinki, 00014, Finland. juha.kere@ki.se.
6
Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, 141 83, Sweden. juha.kere@ki.se.
7
School of Basic and Medical Biosciences, Guy's Hospital, King's College London, London, SE1 9RT, UK. juha.kere@ki.se.
8
Folkhälsan Institute of Genetics, Helsinki, 00290, Finland. juha.kere@ki.se.
9
Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, Helsinki, 00014, Finland. timo.otonkoski@helsinki.fi.
10
Children's Hospital, Helsinki University Central Hospital, University of Helsinki, Helsinki, 00290, Finland. timo.otonkoski@helsinki.fi.

Abstract

CRISPR-Cas9-based gene activation (CRISPRa) is an attractive tool for cellular reprogramming applications due to its high multiplexing capacity and direct targeting of endogenous loci. Here we present the reprogramming of primary human skin fibroblasts into induced pluripotent stem cells (iPSCs) using CRISPRa, targeting endogenous OCT4, SOX2, KLF4, MYC, and LIN28A promoters. The low basal reprogramming efficiency can be improved by an order of magnitude by additionally targeting a conserved Alu-motif enriched near genes involved in embryo genome activation (EEA-motif). This effect is mediated in part by more efficient activation of NANOG and REX1. These data demonstrate that human somatic cells can be reprogrammed into iPSCs using only CRISPRa. Furthermore, the results unravel the involvement of EEA-motif-associated mechanisms in cellular reprogramming.

PMID:
29980666
PMCID:
PMC6035213
DOI:
10.1038/s41467-018-05067-x
[Indexed for MEDLINE]
Free PMC Article

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