Generation and analysis of zebrafish melanoma models.
The rapid emergence of the zebrafish as a cancer model has been aided by advances in genetic, chemical, and imaging technologies. Melanoma in particular highlights both the power and challenges associated with cancer modeling in zebrafish. This chapter focuses on the lessons that have emerged from the melanoma models as paradigmatic of what will apply to nearly all cancer models in the zebrafish system. We specifically focus on methodologies related to germline and mosaic transgenic melanoma generation, and how these can be used to deeply interrogate additional cooperating oncogenes or tumor suppressors. These transgenic tumors can in turn be used to generate zebrafish-specific, stable melanoma cell lines which can be fluorescently labeled, modified by cDNA/CRISPR techniques, and used for detailed in vivo imaging of cancer progression in real time. These zebrafish melanoma models are beginning to elucidate both cell intrinsic and microenvironmental factors in melanoma that have broader implications for human disease. We envision that nearly all of the techniques described here can be applied to other zebrafish cancer models, and likely expanded beyond what we describe here.
MRC Human Genetics Unit, and The University of Edinburgh Cancer Research UK Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, United Kingdom.
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Wojciechowska
Stem Cell Program and the Division of Pediatric Hematology/Oncology, Boston Children's Hospital and Dana-Farber Cancer Institute, Howard Hughes Medical Institute, MA, United States; Harvard Medical School, Boston, MA, United States.
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van Rooijen
Program in Molecular Medicine and Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA, United States.
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Ceol
MRC Human Genetics Unit, and The University of Edinburgh Cancer Research UK Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, United Kingdom.
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Patton
Memorial Sloan Kettering Cancer Center, New York, NY, United States.
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White
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D063646:Carcinogenesis / Q000235:genetics*; D004195:Disease Models, Animal; D006801:Humans; D008545:Melanoma / Q000235:genetics* / Q000473:pathology; D015027:Zebrafish / Q000235:genetics*
Cancer; Cell culture; MITF; Melanoma; Transgenesis; Transplantation; Zebrafish