PT  - JOURNAL ARTICLE
AU  - Maarten H Geurts
AU  - Eyleen de Poel
AU  - Cayetano Pleguezuelos-Manzano
AU  - Rurika Oka
AU  - Léo Carrillo
AU  - Amanda Andersson-Rolf
AU  - Matteo Boretto
AU  - Jesse E Brunsveld
AU  - Ruben van Boxtel
AU  - Jeffrey M Beekman
AU  - Hans Clevers
TI  - Evaluating CRISPR-based prime editing for cancer modeling and CFTR repair in organoids
AID  - 10.26508/lsa.202000940
DP  - 2021 Oct 01
TA  - Life Science Alliance
PG  - e202000940
VI  - 4
IP  - 10
4099  - https://www.life-science-alliance.org/content/4/10/e202000940.short
4100  - https://www.life-science-alliance.org/content/4/10/e202000940.full
SO  - Life Sci. Alliance2021 Oct 01; 4
AB  - 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.