Human pluripotent reprogramming with CRISPR activators.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 29980666.
- Also identified by DOI 10.1038/s41467-018-05067-x and PMC identifier 6035213.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- CRISPR-Associated Protein 9
- Cellular Reprogramming
- Clustered Regularly Interspaced Short Palindromic Repeats
- Induced Pluripotent Stem Cells