In situ CRISPR-Cas9 base editing for the development of genetically engineered mouse models of breast cancer.

TitleIn situ CRISPR-Cas9 base editing for the development of genetically engineered mouse models of breast cancer.
Publication TypeJournal Article
Year of Publication2020
AuthorsAnnunziato S, Lutz C, Henneman L, Bhin J, Wong K, Siteur B, van Gerwen B, de Korte-Grimmerink R, Zafra MPaz, Schatoff EM, Drenth APaulien, van der Burg E, Eijkman T, Mukherjee S, Boroviak K, Wessels LFa, van de Ven M, Huijbers IJ, Adams DJ, Dow LE, Jonkers J
JournalEMBO J
Paginatione102169
Date Published2020 Jan 13
ISSN1460-2075
Abstract

Genetically engineered mouse models (GEMMs) of cancer have proven to be of great value for basic and translational research. Although CRISPR-based gene disruption offers a fast-track approach for perturbing gene function and circumvents certain limitations of standard GEMM development, it does not provide a flexible platform for recapitulating clinically relevant missense mutations in vivo. To this end, we generated knock-in mice with Cre-conditional expression of a cytidine base editor and tested their utility for precise somatic engineering of missense mutations in key cancer drivers. Upon intraductal delivery of sgRNA-encoding vectors, we could install point mutations with high efficiency in one or multiple endogenous genes in situ and assess the effect of defined allelic variants on mammary tumorigenesis. While the system also produces bystander insertions and deletions that can stochastically be selected for when targeting a tumor suppressor gene, we could effectively recapitulate oncogenic nonsense mutations. We successfully applied this system in a model of triple-negative breast cancer, providing the proof of concept for extending this flexible somatic base editing platform to other tissues and tumor types.

DOI10.15252/embj.2019102169
Alternate JournalEMBO J.
PubMed ID31930530
Grant ListVICI 91814643 / / Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO) /
024001028 / / Cancer Genomics Netherlands (CGCNL) /
85300120 / / Cancer Systems Biology Center (CSBC) /
93512009 / / Netherlands Genomics Initiative (NGI) Zenith /
/ / EC | FP7 | FP7 Ideas: European Research Council (FP7 Ideas) /
319661 / / ERC Synergy project CombatCancer /
184032303 / / National Roadmap grant for Large-Scale Research Facilities /
/ / Oncode Institute /

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