Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells Journal Article

   


Authors: Wu, Y.; Zhong, A.; Sidharta, M.; Kim, T. W.; Ramirez, B.; Persily, B.; Studer, L.; Zhou, T.
Article Title: Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells
Abstract: Prime editing (PE) allows for precise genome editing in human pluripotent stem cells (hPSCs), such as introducing single nucleotide modifications, small insertions or deletions at a specific genomic locus. Here, we systematically compare a panel of prime editing conditions in hPSCs and generate a potent prime editor, “PE-Plus”, through co-inhibition of mismatch repair and p53-mediated cellular stress responses. We further establish an inducible prime editing platform in hPSCs by incorporating the PE-Plus into a safe-harbor locus and demonstrated temporal control of precise editing in both hPSCs and differentiated cells. By evaluating disease-associated mutations, we show that this platform allows efficient creation of both monoallelic and biallelic disease-relevant mutations in hPSCs. In addition, this platform enables the efficient introduction of single or multiple edits in one step, demonstrating potential for multiplex editing. Our method presents an efficient and controllable multiplex prime editing tool in hPSCs and their differentiated progeny. © The Author(s) 2024.
Keywords: controlled study; gene mutation; human cell; frameshift mutation; genetics; mutation; stop codon; nonhuman; cytology; metabolism; allele; gene expression; carboxy terminal sequence; embryo; epidermal growth factor receptor; cell line; gene locus; gene function; cell differentiation; cell population; protein p53; amino terminal sequence; mismatch repair; human genome; plasmid; lentivirus; tumor suppressor protein p53; electroporation; genome; pluripotent stem cell; pluripotent stem cells; mutation rate; cell clone; dna binding; doxycycline; cellular stress response; nuclear localization signal; genome, human; cell; genotyping; neuroectoderm; procedures; indel mutation; disease incidence; off-target effect; sanger sequencing; humans; human; article; whole genome sequencing; crispr cas system; induced mutation; droplet digital polymerase chain reaction; gene editing; crispr-cas systems; hek293t cell line; human embryonic stem cell; prime editing; tetramerization
Journal Title: Nature Communications
Volume: 15
ISSN: 2041-1723
Publisher: Nature Publishing Group  
Date Published: 2024-12-30
Start Page: 10824
Language: English
DOI: 10.1038/s41467-024-55104-1
PUBMED: 39737975
PROVIDER: scopus
PMCID: PMC11685797
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85214013494&doi=10.1038%2fs41467-024-55104-1&partnerID=40&md5=8b38b890d24603bb3ce95d443fea29d5
Notes: The MSK Cancer Center Support Grant (P30 CA008748) is acknowledged in the PubMed record and PDF. Corresponding MSK authors are Lorenz Studer and Ting Zhou -- Source: Scopus
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MSK Authors
  1. Lorenz Studer
    223 Studer
  2. Taewan Kim
    11 Kim
  3. Ting Zhou
    29 Zhou
  4. Aaron Zhong
    11 Zhong
  5. Mega Sidharta
    5 Sidharta
  6. Benjamin Scott Persily
    2 Persily
  7. Youjun Wu
    4 Wu
  8. Bernny Jose Ramirez
    1 Ramirez
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