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JoVE Journal Cancer Research
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In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System

Article DOI: 10.3791/60410-v • 05:45 min • January 9th, 2020
January 9th, 2020 •
*1,2, *1,2, 3, 2,4, 1,2
1The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, 2Faculty of Health Sciences, Ben-Gurion University of the Negev, 3Human Oncology & Pathogenesis Program (HOPP), Memorial Sloan Kettering Cancer Center, 4Institute of Pathology, Barzilai University Medical Center
* These authors contributed equally

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Summary
Automatic Translation

Development of murine models with specific genes mutated in head and neck cancer patients is required for understanding of neoplasia. Here, we present a protocol for in vitro transformation of primary murine tongue cells using an adeno-associated virus-Cas9 system to generate murine HNC cell lines with specific genomic alterations.

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In Vitro Genetically Engineered Murine Head And Neck Cancer Cell Line Adeno-associated Virus-Cas9 System Protocol Genomic Operation Neoplasia Specific Gene Mutations Organ Surgical Scissors Harvest Tongue B6 Transgenic Mouse Scalpel Tissue Fragments RPMI Plain Medium Without Serum Triple Enzyme Mix Incubation Enzymatic Dissociation 5%FBS In PBS Cell Suspension 70 Micron Mesh Strainer Centrifugation Complete Medium Cell Aggregates
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