Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells Journal Article


Authors: Li, M.; Zhong, A.; Wu, Y.; Sidharta, M.; Beaury, M.; Zhao, X.; Studer, L.; Zhou, T.
Article Title: Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells
Abstract: Precise gene editing in human pluripotent stem cells (hPSCs) holds great promise for studying and potentially treating human diseases. Both prime editing and base editing avoid introducing double strand breaks, but low editing efficiencies make those techniques still an arduous process in hPSCs. Here we report that co-delivering of p53DD, a dominant negative fragment of p53, can greatly enhance prime editing and cytosine base editing efficiencies in generating precise mutations in hPSCs. We further apply PE3 in combination with p53DD to efficiently create multiple isogenic hPSC lines, including lines carrying GBA or LRRK2 mutations associated with Parkinson disease and a LMNA mutation linked to Hutchinson-Gilford progeria syndrome. We also correct GBA and LMNA mutations in the patient-specific iPSCs. Our data show that p53DD improves PE3 efficiency without compromising the genome-wide safety, making it feasible for safe and routine generation of isogenic hPSC lines for disease modeling. © 2022, The Author(s).
Keywords: controlled study; unclassified drug; human cell; promoter region; single nucleotide polymorphism; genetics; mutation; stop codon; flow cytometry; gene expression; cell protein; small interfering rna; gene locus; transcriptomics; protein p53; physiology; deamination; ubiquitination; cytidine deaminase; plasmid; tumor suppressor protein p53; electroporation; pluripotent stem cell; pluripotent stem cells; cyclin dependent kinase inhibitor 1a; cell clone; point mutation; parkinson disease; fluorescence activated cell sorting; transcriptome; genomic dna; induced pluripotent stem cells; expression vector; start codon; inhibition; cell; cytosine; procedures; indel mutation; leucine rich repeat kinase 2; gene expression system; induced pluripotent stem cell; sanger sequencing; humans; human; article; whole genome sequencing; rna sequencing; pluripotent stem cell line; crispr cas system; gene editing; crispr-cas systems; progeria; human embryonic stem cell; gba protein; p53dd protein; pe3 protein; reading frame
Journal Title: Nature Communications
Volume: 13
ISSN: 2041-1723
Publisher: Nature Publishing Group  
Date Published: 2022-10-27
Start Page: 6354
Language: English
DOI: 10.1038/s41467-022-34045-7
PUBMED: 36302757
PROVIDER: scopus
PMCID: PMC9613702
DOI/URL:
Notes: Article -- Export Date: 1 December 2022 -- Source: Scopus
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Citation Impact
MSK Authors
  1. Xiaolan Zhao
    73 Zhao
  2. Lorenz Studer
    195 Studer
  3. Ting Zhou
    13 Zhou
  4. Aaron Zhong
    8 Zhong
  5. Mu Li
    2 Li
  6. Michael Beaury
    1 Beaury