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Human pluripotent reprogramming with CRISPR activators.

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.

SUBMITTER: Weltner J

PROVIDER: S-EPMC6035213 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Human pluripotent reprogramming with CRISPR activators.

Weltner Jere J Balboa Diego D Katayama Shintaro S Bespalov Maxim M Krjutškov Kaarel K Jouhilahti Eeva-Mari EM Trokovic Ras R Kere Juha J Otonkoski Timo T

Nature communications 20180706 1

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- ...[more]

PMID: 29980666

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