Cancer modeling by Transgene Electroporation in Adult Zebrafish (TEAZ)

被引:37
作者
Callahan, Scott J. [1 ,2 ,3 ,4 ]
Tepan, Stephanie [5 ,6 ]
Zhang, Yan M. [1 ,2 ]
Lindsay, Helen [7 ,8 ]
Burger, Alexa [7 ]
Campbell, Nathaniel R. [9 ]
Kim, Isabella S. [1 ,2 ]
Hollmann, Travis J. [10 ]
Studer, Lorenz [11 ,12 ]
Mosimann, Christian [7 ]
White, Richard M. [1 ,2 ,13 ]
机构
[1] Mem Sloan Kettering Canc Ctr, Canc Biol & Genet, New York, NY 10065 USA
[2] Dept Med, New York, NY 10065 USA
[3] Mem Sloan Kettering Canc Ctr, Dev Biol, New York, NY 10065 USA
[4] Mem Sloan Kettering Canc Ctr, Gerstner Grad Sch Biomed Sci, New York, NY 10065 USA
[5] Mem Sloan Kettering Canc Ctr, Summer Clin Oncol Res Experience SCORE Program 20, New York, NY 10065 USA
[6] Hunter Coll, New York, NY 10065 USA
[7] Univ Zurich, Inst Mol Life Sci, CH-8057 Zurich, Switzerland
[8] Univ Zurich, SIB, CH-8057 Zurich, Switzerland
[9] Weill Cornell Rockefeller Sloan Kettering Triinst, New York, NY 10065 USA
[10] Mem Sloan Kettering Canc Ctr, Pathol, New York, NY 10065 USA
[11] Sloan Kettering Inst, Ctr Stem Cell Biol, New York, NY 10065 USA
[12] Sloan Kettering Inst, Dev Biol Program, New York, NY 10065 USA
[13] Weill Cornell Med Coll, New York, NY 10065 USA
基金
美国国家卫生研究院; 瑞士国家科学基金会;
关键词
Cancer; Electroporation; Melanoma; Zebrafish; Metastasis; IN-VIVO ELECTROPORATION; SOFT-TISSUE SARCOMAS; NEURAL-CREST; WEB TOOL; GENE; TRANSPOSON; P53; EXPRESSION; MUTATIONS; INACTIVATION;
D O I
10.1242/dmm.034561
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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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页数:10
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