Engraftment of Allotransplanted Tumor Cells in Adult Mutant .

Modeling human genetic diseases and cancer in lab animals has been greatly aided by the emergence of genetic engineering tools such as TALENs and CRISPR/Cas9. We have previously demonstrated the ease with which genetically engineered models (GEXM) can be generated via injection of early embryos with Cas9 recombinant protein loaded with sgRNAs targeting single or multiple tumor suppressor genes. What has been lacking so far is the possibility to propagate and characterize the induced cancers via transplantation. Here, we describe the generation of a knockout line in that is deficient in functional T and B cells. This line was validated by means of allografting experiments with primary and donor tumors. In addition, we optimized available protocols for the sub-lethal irradiation of wild-type froglets. Irradiated animals also allowed the stable, albeit transient, engraftment of transplanted tumor cells. The novel line and the irradiated wild-type froglets will further expand the experimental toolbox in the diploid amphibian and help to establish it as a versatile and relevant model for exploring human cancer.