Abstract
Transgenic expression of genes is a mainstay of cancer modeling in zebrafish. Traditional transgenic techniques rely upon injection into one-cell embryos, but ideally these transgenes would be expressed only in adult somatic tissues. We provide a method to model cancer in adult zebrafish in which transgenes can be expressed via electroporation. Using melanoma as an example, we demonstrate the feasibility of expressing oncogenes such as BRAFV600E as well as CRISPR/Cas9 inactivation of tumor suppressors such as PTEN. These approaches can be performed in any genetic background such as existing fluorophore reporter lines or the casper line. These methods can readily be extended to other cell types allowing for rapid adult modeling of cancer in zebrafish.
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References
Stuart GW, McMurray JV, Westerfield M (1988) Replication, integration and stable germ-line transmission of foreign sequences injected into early zebrafish embryos. Development 103:403–412
Lawson ND, Weinstein BM (2002) In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev Biol 248:307–318. https://doi.org/10.1006/dbio.2002.0711
Long Q, Meng A, Wang H et al (1997) GATA-1 expression pattern can be recapitulated in living transgenic zebrafish using GFP reporter gene. Development 124:4105–4111
Lister JA, Robertson CP, Lepage T et al (1999) Nacre encodes a zebrafish microphthalmia-related protein that regulates neural-crest-derived pigment cell fate. Development 126:3757–3767
Langenau DM, Traver D, Ferrando AA et al (2003) Myc-induced T cell leukemia in transgenic zebrafish. Science 299:887–890. https://doi.org/10.1126/science.1080280
Patton EE, Widlund HR, Kutok JL et al (2005) BRAF mutations are sufficient to promote nevi formation and cooperate with p53 in the genesis of melanoma. Curr Biol 15:249–254. https://doi.org/10.1016/j.cub.2005.01.031
Ablain J, Xu M, Rothschild H et al (2018) Human tumor genomics and zebrafish modeling identify SPRED1 loss as a driver of mucosal melanoma. Science 362:1055–1060. https://doi.org/10.1126/science.aau6509
Langenau DM, Keefe MD, Storer NY et al (2008) Co-injection strategies to modify radiation sensitivity and tumor initiation in transgenic zebrafish. Oncogene 27:4242–4248. https://doi.org/10.1038/onc.2008.56
Nguyen AT, Koh V, Spitsbergen JM, Gong Z (2016) Development of a conditional liver tumor model by mifepristone-inducible Cre recombination to control oncogenic kras V12 expression in transgenic zebrafish. Sci Rep 6:19559. https://doi.org/10.1038/srep19559
Langenau DM, Feng H, Berghmans S et al (2005) Cre/lox-regulated transgenic zebrafish model with conditional myc-induced T cell acute lymphoblastic leukemia. Proc Natl Acad Sci U S A 102:6068–6073. https://doi.org/10.1073/pnas.0408708102
Callahan SJ, Tepan S, Zhang YM et al (2018) Cancer modeling by transgene electroporation in adult zebrafish (TEAZ). Dis Model Mech 11. https://doi.org/10.1242/dmm.034561
White RM, Sessa A, Burke C et al (2008) Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell 2:183–189. https://doi.org/10.1016/j.stem.2007.11.002
Heilmann S, Ratnakumar K, Langdon E et al (2015) A quantitative system for studying metastasis using transparent zebrafish. Cancer Res 75:4272–4282. https://doi.org/10.1158/0008-5472.CAN-14-3319
Kwan KM, Fujimoto E, Grabher C et al (2007) The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev Dyn 236:3088–3099. https://doi.org/10.1002/dvdy.21343
Yin L, Maddison LA, Li M et al (2015) Multiplex conditional mutagenesis using transgenic expression of Cas9 and sgRNAs. Genetics 200:431–441. https://doi.org/10.1534/genetics.115.176917
Ceol CJ, Houvras Y, Jane-Valbuena J et al (2011) The histone methyltransferase SETDB1 is recurrently amplified in melanoma and accelerates its onset. Nature 471:513–517. https://doi.org/10.1038/nature09806
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Montal, E., Suresh, S., Ma, Y., Tagore, M.M., White, R.M. (2024). Cancer Modeling by Transgene Electroporation in Adult Zebrafish (TEAZ). In: Amatruda, J.F., Houart, C., Kawakami, K., Poss, K.D. (eds) Zebrafish. Methods in Molecular Biology, vol 2707. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3401-1_5
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DOI: https://doi.org/10.1007/978-1-0716-3401-1_5
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