RT Journal Article
SR Electronic
T1 Evaluating CRISPR-based prime editing for cancer modeling and CFTR repair in organoids
JF Life Science Alliance
JO Life Sci. Alliance
FD Life Science Alliance LLC
SP e202000940
DO 10.26508/lsa.202000940
VO 4
IS 10
A1 Maarten H Geurts
A1 Eyleen de Poel
A1 Cayetano Pleguezuelos-Manzano
A1 Rurika Oka
A1 Léo Carrillo
A1 Amanda Andersson-Rolf
A1 Matteo Boretto
A1 Jesse E Brunsveld
A1 Ruben van Boxtel
A1 Jeffrey M Beekman
A1 Hans Clevers
YR 2021
UL https://www.life-science-alliance.org/content/4/10/e202000940.abstract
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.