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One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids

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 c...

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Autores principales: Geurts, Maarten H., Gandhi, Shashank, Boretto, Matteo G., Akkerman, Ninouk, Derks, Lucca L. M., van Son, Gijs, Celotti, Martina, Harshuk-Shabso, Sarina, Peci, Flavia, Begthel, Harry, Hendriks, Delilah, Schürmann, Paul, Andersson-Rolf, Amanda, Chuva de Sousa Lopes, Susana M., van Es, Johan H., van Boxtel, Ruben, Clevers, Hans
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10435570/
https://www.ncbi.nlm.nih.gov/pubmed/37591832
http://dx.doi.org/10.1038/s41467-023-40701-3
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author Geurts, Maarten H.
Gandhi, Shashank
Boretto, Matteo G.
Akkerman, Ninouk
Derks, Lucca L. M.
van Son, Gijs
Celotti, Martina
Harshuk-Shabso, Sarina
Peci, Flavia
Begthel, Harry
Hendriks, Delilah
Schürmann, Paul
Andersson-Rolf, Amanda
Chuva de Sousa Lopes, Susana M.
van Es, Johan H.
van Boxtel, Ruben
Clevers, Hans
author_facet Geurts, Maarten H.
Gandhi, Shashank
Boretto, Matteo G.
Akkerman, Ninouk
Derks, Lucca L. M.
van Son, Gijs
Celotti, Martina
Harshuk-Shabso, Sarina
Peci, Flavia
Begthel, Harry
Hendriks, Delilah
Schürmann, Paul
Andersson-Rolf, Amanda
Chuva de Sousa Lopes, Susana M.
van Es, Johan H.
van Boxtel, Ruben
Clevers, Hans
author_sort Geurts, Maarten H.
collection PubMed
description 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.
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spelling pubmed-104355702023-08-19 One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids Geurts, Maarten H. Gandhi, Shashank Boretto, Matteo G. Akkerman, Ninouk Derks, Lucca L. M. van Son, Gijs Celotti, Martina Harshuk-Shabso, Sarina Peci, Flavia Begthel, Harry Hendriks, Delilah Schürmann, Paul Andersson-Rolf, Amanda Chuva de Sousa Lopes, Susana M. van Es, Johan H. van Boxtel, Ruben Clevers, Hans Nat Commun Article 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. Nature Publishing Group UK 2023-08-17 /pmc/articles/PMC10435570/ /pubmed/37591832 http://dx.doi.org/10.1038/s41467-023-40701-3 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Geurts, Maarten H.
Gandhi, Shashank
Boretto, Matteo G.
Akkerman, Ninouk
Derks, Lucca L. M.
van Son, Gijs
Celotti, Martina
Harshuk-Shabso, Sarina
Peci, Flavia
Begthel, Harry
Hendriks, Delilah
Schürmann, Paul
Andersson-Rolf, Amanda
Chuva de Sousa Lopes, Susana M.
van Es, Johan H.
van Boxtel, Ruben
Clevers, Hans
One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
title One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
title_full One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
title_fullStr One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
title_full_unstemmed One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
title_short One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
title_sort one-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10435570/
https://www.ncbi.nlm.nih.gov/pubmed/37591832
http://dx.doi.org/10.1038/s41467-023-40701-3
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