TY - JOUR
T1 - One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
AU - Geurts, Maarten H.
AU - Gandhi, Shashank
AU - Boretto, Matteo G.
AU - Akkerman, Ninouk
AU - Derks, Lucca L.M.
AU - van Son, Gijs
AU - Celotti, Martina
AU - Harshuk-Shabso, Sarina
AU - Peci, Flavia
AU - Begthel, Harry
AU - Hendriks, Delilah
AU - Schürmann, Paul
AU - Andersson-Rolf, Amanda
AU - Chuva de Sousa Lopes, Susana M.
AU - van Es, Johan H.
AU - van Boxtel, Ruben
AU - Clevers, Hans
N1 - Publisher Copyright:
© 2023, Springer Nature Limited.
PY - 2023/8/17
Y1 - 2023/8/17
N2 - Optimization of CRISPR/Cas9-mediated genome engineering has resulted in base editors that hold promise for mutation repair and disease modeling. Here, we demonstrate the application of base editors for the generation of complex tumor models in human ASC-derived organoids. First we show efficacy of cytosine and adenine base editors in modeling CTNNB1 hot-spot mutations in hepatocyte organoids. Next, we use C > T base editors to insert nonsense mutations in PTEN in endometrial organoids and demonstrate tumorigenicity even in the heterozygous state. Moreover, drug sensitivity assays on organoids harboring either PTEN or PTEN and PIK3CA mutations reveal the mechanism underlying the initial stages of endometrial tumorigenesis. To further increase the scope of base editing we combine SpCas9 and SaCas9 for simultaneous C > T and A > G editing at individual target sites. Finally, we show that base editor multiplexing allow modeling of colorectal tumorigenesis in a single step by simultaneously transfecting sgRNAs targeting five cancer genes.
AB - Optimization of CRISPR/Cas9-mediated genome engineering has resulted in base editors that hold promise for mutation repair and disease modeling. Here, we demonstrate the application of base editors for the generation of complex tumor models in human ASC-derived organoids. First we show efficacy of cytosine and adenine base editors in modeling CTNNB1 hot-spot mutations in hepatocyte organoids. Next, we use C > T base editors to insert nonsense mutations in PTEN in endometrial organoids and demonstrate tumorigenicity even in the heterozygous state. Moreover, drug sensitivity assays on organoids harboring either PTEN or PTEN and PIK3CA mutations reveal the mechanism underlying the initial stages of endometrial tumorigenesis. To further increase the scope of base editing we combine SpCas9 and SaCas9 for simultaneous C > T and A > G editing at individual target sites. Finally, we show that base editor multiplexing allow modeling of colorectal tumorigenesis in a single step by simultaneously transfecting sgRNAs targeting five cancer genes.
UR - http://www.scopus.com/inward/record.url?scp=85168274131&partnerID=8YFLogxK
U2 - 10.1038/s41467-023-40701-3
DO - 10.1038/s41467-023-40701-3
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C2 - 37591832
AN - SCOPUS:85168274131
SN - 2041-1723
VL - 14
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 4998
ER -
