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Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model
Published on: November 13, 2012
Screening for melanoma modifiers using a zebrafish autochthonous tumor model
Sharanya Iyengar1, Yariv Houvras, Craig J Ceol
1Program in Molecular Medicine and Department of Cancer Biology, University of Massachusetts Medical School, USA.
Journal of Visualized Experiments : Jove
|November 28, 2012
Summary
This study introduces a rapid zebrafish model to screen melanoma-driving genes. The miniCoopR vector enables quantitative assessment of candidate gene effects on melanoma initiation and progression.
Area of Science:
- Genetics
- Cancer Research
- Developmental Biology
Background:
- Genomic studies reveal numerous gene alterations in human cancers.
- Distinguishing driver mutations from passenger mutations in tumorigenesis is challenging.
- A quantitative assay is needed to assess the impact of altered genes on tumor development.
Purpose of the Study:
- To develop a rapid zebrafish assay for screening candidate melanoma modifiers.
- To quantitatively measure the effect of altered genes on melanoma initiation and persistence.
Main Methods:
- Utilized an autochthonous zebrafish tumor model for melanoma initiation and maintenance.
- Employed the miniCoopR vector, coupling a wild-type mitfa minigene with a Gateway recombination cassette.
- Injected constructs into Tg(mitfa:BRAF(V600E));p53(lf);mitfa(lf) zebrafish embryos for Tol2-mediated transgenesis.
Main Results:
- The miniCoopR vector facilitates expression of candidate genes in rescued melanocytes.
- Transformed melanocytes developed into tumors, enabling melanoma initiation studies.
- Melanoma-free survival, invasion, and transplantation assays quantified gene effects.
Conclusions:
- The developed zebrafish model provides a rapid and quantitative method for screening melanoma modifiers.
- This assay aids in distinguishing genes driving tumorigenesis from those arising incidentally.
- The miniCoopR vector is a key tool for functional genomics in melanoma research.
