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

Juho Joonas Sokka, Masahito Yoshihara, Jouni Kvist, Laura Laiho, Andrew Warren, Christian Stadelmann, Eeva-Mari Jouhilahti, Helena Kilpinen, Diego Balboa, Shintaro Katayama, Aija Kyttälä, Juha Kere, Timo Otonkoski, Jere Weltner, Ras Trokovic

Research output: Contribution to journalArticleScientificpeer-review


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.
Original languageEnglish
JournalStem cell reports
Issue number2
Pages (from-to)413–426
Number of pages14
Publication statusPublished - 8 Feb 2022
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 1184 Genetics, developmental biology, physiology
  • Reprogramming
  • ingle cell RNA sequencing
  • CRISPR-Cas9
  • Transcriptomics
  • 1182 Biochemistry, cell and molecular biology
  • MicroRNA (miRNA)
  • iPSC
  • Stem cells
  • GENE

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