Cancer Biology and Genetics, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Methods Mol Biol. 2024;2707:83-97. doi: 10.1007/978-1-0716-3401-1_5.
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.
转基因表达是斑马鱼癌症建模的主要方法。传统的转基因技术依赖于注射到单细胞胚胎中,但理想情况下,这些转基因只能在成年体细胞组织中表达。我们提供了一种通过电穿孔在成年斑马鱼中建模癌症的方法,其中转基因可以表达。我们以黑色素瘤为例,证明了表达癌基因如 BRAFV600E 以及 CRISPR/Cas9 失活肿瘤抑制基因如 PTEN 的可行性。这些方法可以在任何遗传背景下进行,如现有的荧光报告基因系或 casper 系。这些方法可以很容易地扩展到其他细胞类型,从而可以快速在斑马鱼中进行癌症的成年建模。