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In situ CRISPR-Cas9 base editing for the development of genetically engineered mouse models of breast cancer.
Annunziato, Stefano; Lutz, Catrin; Henneman, Linda; Bhin, Jinhyuk; Wong, Kim; Siteur, Bjørn; van Gerwen, Bas; de Korte-Grimmerink, Renske; Zafra, Maria Paz; Schatoff, Emma M; Drenth, Anne Paulien; van der Burg, Eline; Eijkman, Timo; Mukherjee, Siddhartha; Boroviak, Katharina; Wessels, Lodewyk Fa; van de Ven, Marieke; Huijbers, Ivo J; Adams, David J; Dow, Lukas E; Jonkers, Jos.
Afiliação
  • Annunziato S; Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Lutz C; Cancer Genomics Netherlands, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Henneman L; Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Bhin J; Cancer Genomics Netherlands, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Wong K; Transgenic Core Facility, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Siteur B; Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • van Gerwen B; Cancer Genomics Netherlands, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • de Korte-Grimmerink R; Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Zafra MP; Wellcome Trust Sanger Institute, Cambridge, UK.
  • Schatoff EM; Preclinical Intervention Unit, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Drenth AP; Preclinical Intervention Unit, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • van der Burg E; Preclinical Intervention Unit, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Eijkman T; Division of Hematology and Medical Oncology, Department of Medicine, Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA.
  • Mukherjee S; Division of Hematology and Medical Oncology, Department of Medicine, Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA.
  • Boroviak K; Weill Cornell/Rockefeller/Sloan Kettering Tri-I MD-PhD Program, New York, NY, USA.
  • Wessels LF; Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • van de Ven M; Cancer Genomics Netherlands, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Huijbers IJ; Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Adams DJ; Cancer Genomics Netherlands, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Dow LE; Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Jonkers J; Cancer Genomics Netherlands, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
EMBO J ; 39(5): e102169, 2020 03 02.
Article em En | MEDLINE | ID: mdl-31930530
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.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Neoplasias da Mama / Sistemas CRISPR-Cas / Edição de Genes Limite: Animals Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Neoplasias da Mama / Sistemas CRISPR-Cas / Edição de Genes Limite: Animals Idioma: En Ano de publicação: 2020 Tipo de documento: Article