Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i>
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 <i>Xenopus</i> models (GEXM) can be generated via inje...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/19/4560 |
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| author | Dieter Tulkens Dionysia Dimitrakopoulou Marthe Boelens Tom Van Nieuwenhuysen Suzan Demuynck Wendy Toussaint David Creytens Pieter Van Vlierberghe Kris Vleminckx |
| author_facet | Dieter Tulkens Dionysia Dimitrakopoulou Marthe Boelens Tom Van Nieuwenhuysen Suzan Demuynck Wendy Toussaint David Creytens Pieter Van Vlierberghe Kris Vleminckx |
| author_sort | Dieter Tulkens |
| collection | DOAJ |
| description | 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 <i>Xenopus</i> 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 <i>rag2</i> knockout line in <i>Xenopus tropicalis</i> that is deficient in functional T and B cells. This line was validated by means of allografting experiments with primary <i>tp53</i><sup>−/−</sup> and <i>apc<sup>+/−</sup>/tp53<sup>−/−</sup></i> donor tumors. In addition, we optimized available protocols for the sub-lethal irradiation of wild-type <i>X. tropicalis</i> froglets. Irradiated animals also allowed the stable, albeit transient, engraftment of transplanted <i>X. tropicalis</i> tumor cells. The novel <i>rag2</i><sup>−/−</sup> line and the irradiated wild-type froglets will further expand the experimental toolbox in the diploid amphibian <i>X. tropicalis</i> and help to establish it as a versatile and relevant model for exploring human cancer. |
| first_indexed | 2024-03-09T21:58:23Z |
| format | Article |
| id | doaj.art-2b96dff93ca44142a0b58ddb2e3eeba4 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-09T21:58:23Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-2b96dff93ca44142a0b58ddb2e3eeba42023-11-23T19:52:49ZengMDPI AGCancers2072-66942022-09-011419456010.3390/cancers14194560Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i>Dieter Tulkens0Dionysia Dimitrakopoulou1Marthe Boelens2Tom Van Nieuwenhuysen3Suzan Demuynck4Wendy Toussaint5David Creytens6Pieter Van Vlierberghe7Kris Vleminckx8Department of Biomedical Molecular Biology, Ghent University, 9000 Ghent, BelgiumDepartment of Biomedical Molecular Biology, Ghent University, 9000 Ghent, BelgiumDepartment of Biomedical Molecular Biology, Ghent University, 9000 Ghent, BelgiumDepartment of Biomedical Molecular Biology, Ghent University, 9000 Ghent, BelgiumDepartment of Biomedical Molecular Biology, Ghent University, 9000 Ghent, BelgiumDepartment of Internal Medicine and Pediatrics, Ghent University, 9000 Ghent, BelgiumCancer Research Institute Ghent (CRIG), 9000 Ghent, BelgiumCancer Research Institute Ghent (CRIG), 9000 Ghent, BelgiumDepartment of Biomedical Molecular Biology, Ghent University, 9000 Ghent, BelgiumModeling 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 <i>Xenopus</i> 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 <i>rag2</i> knockout line in <i>Xenopus tropicalis</i> that is deficient in functional T and B cells. This line was validated by means of allografting experiments with primary <i>tp53</i><sup>−/−</sup> and <i>apc<sup>+/−</sup>/tp53<sup>−/−</sup></i> donor tumors. In addition, we optimized available protocols for the sub-lethal irradiation of wild-type <i>X. tropicalis</i> froglets. Irradiated animals also allowed the stable, albeit transient, engraftment of transplanted <i>X. tropicalis</i> tumor cells. The novel <i>rag2</i><sup>−/−</sup> line and the irradiated wild-type froglets will further expand the experimental toolbox in the diploid amphibian <i>X. tropicalis</i> and help to establish it as a versatile and relevant model for exploring human cancer.https://www.mdpi.com/2072-6694/14/19/4560<i>Xenopus</i>tumor transplantationsgenetic cancer model<i>rag2</i> knockoutimmunocompromised |
| spellingShingle | Dieter Tulkens Dionysia Dimitrakopoulou Marthe Boelens Tom Van Nieuwenhuysen Suzan Demuynck Wendy Toussaint David Creytens Pieter Van Vlierberghe Kris Vleminckx Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i> Cancers <i>Xenopus</i> tumor transplantations genetic cancer model <i>rag2</i> knockout immunocompromised |
| title | Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i> |
| title_full | Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i> |
| title_fullStr | Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i> |
| title_full_unstemmed | Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i> |
| title_short | Engraftment of Allotransplanted Tumor Cells in Adult <i>rag2</i> Mutant <i>Xenopus tropicalis</i> |
| title_sort | engraftment of allotransplanted tumor cells in adult i rag2 i mutant i xenopus tropicalis i |
| topic | <i>Xenopus</i> tumor transplantations genetic cancer model <i>rag2</i> knockout immunocompromised |
| url | https://www.mdpi.com/2072-6694/14/19/4560 |
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