期刊
DISEASE MODELS & MECHANISMS
卷 11, 期 9, 页码 -出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/dmm.034561
关键词
Cancer; Electroporation; Melanoma; Zebrafish; Metastasis
资金
- National Institutes of Health [DP2CA186572, P30CA008748, K08AR055368]
- National Cancer Institute [F31CA196305, 1F99CA212436-01]
- Melanoma Research Alliance
- Starr Cancer Consortium
- Pershing Square Sohn Foundation
- Harry J. Lloyd Foundation
- Consano
- Joanna M. Nicolay Melanoma Foundation
- Schweizerischer Nationalfonds zur Forderung der Wissenschaftlichen Forschung [PP00P3_139093]
- SwissBridge Foundation
- FP7 People: Marie-Curie Actions [CIG PCIG14-GA-2013-631984]
- National Institute of General Medical Sciences [T32GM007739]
- Melanoma Research Foundation
- Alan and Sandra Gerry Metastasis and Tumor Ecosystems Center at Memorial Sloan Kettering Cancer Center
- Robert B. Catell Fellowship at Memorial Sloan Kettering Cancer Center
- NATIONAL CANCER INSTITUTE [F31CA196305, P30CA008748, F99CA212436, DP2CA186572, F30CA220954] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES [K08AR055368] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [T32GM007739] Funding Source: NIH RePORTER
Transgenic animals are invaluable for modeling cancer genomics, but often require complex crosses of multiple germline alleles to obtain the desired combinations. Zebrafish models have advantages in that transgenes can be rapidly tested by mosaic expression, but typically lack spatial and temporal control of tumor onset, which limits their utility for the study of tumor progression and metastasis. To overcome these limitations, we have developed a method referred to as Transgene Electroporation in Adult Zebrafish (TEAZ). TEAZ can deliver DNA constructs with promoter elements of interest to drive fluorophores, oncogenes or CRISPR-Cas9-based mutagenic cassettes in specific cell types. Using TEAZ, we created a highly aggressive melanoma model via Cas9-mediated inactivation of Rb1 in the context of BRAF(V600E) in spatially constrained melanocytes. Unlike prior models that take similar to 4 months to develop, we found that TEAZ leads to tumor onset in similar to 7 weeks, and these tumors develop in fully immunocompetent animals. As the resulting tumors initiated at highly defined locations, we could track their progression via fluorescence, and documented deep invasion into tissues and metastatic deposits. TEAZ can be deployed to other tissues and cell types, such as the heart, with the use of suitable transgenic promoters. The versatility of TEAZ makes it widely accessible for rapid modeling of somatic gene alterations and cancer progression at a scale not achievable in other in vivo systems.
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