Evaluating CRISPR-based prime editing for cancer modeling and CFTR repair in organoids

Maarten H Geurts, Eyleen de Poel, Cayetano Pleguezuelos-Manzano, Rurika Oka, Léo Carrillo, Amanda Andersson-Rolf, Matteo Boretto, Jesse E Brunsveld, Ruben van Boxtel, Jeffrey M Beekman, Hans Clevers

Research output: Contribution to journalArticlepeer-review

49 Citations (Scopus)


Prime editing is a recently reported genome editing tool using a nickase-cas9 fused to a reverse transcriptase that directly synthesizes the desired edit at the target site. Here, we explore the use of prime editing in human organoids. Common TP53 mutations can be correctly modeled in human adult stem cell-derived colonic organoids with efficiencies up to 25% and up to 97% in hepatocyte organoids. Next, we functionally repaired the cystic fibrosis CFTR-F508del mutation and compared prime editing to CRISPR/Cas9-mediated homology-directed repair and adenine base editing on the CFTR-R785* mutation. Whole-genome sequencing of prime editing-repaired organoids revealed no detectable off-target effects. Despite encountering varying editing efficiencies and undesired mutations at the target site, these results underline the broad applicability of prime editing for modeling oncogenic mutations and showcase the potential clinical application of this technique, pending further optimization.

Original languageEnglish
Article numbere202000940
JournalLife science alliance
Issue number10
Publication statusPublished - Oct 2021


  • Alleles
  • Amino Acid Substitution
  • Animals
  • CRISPR-Cas Systems
  • Cystic Fibrosis Transmembrane Conductance Regulator/genetics
  • Epithelial Cells/metabolism
  • Gene Editing
  • Genetic Therapy/methods
  • Hepatocytes/metabolism
  • Humans
  • Mutation
  • Oncogenes/genetics
  • Organoids
  • Sequence Analysis, DNA


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