Use of Cre-adenovirus and CAR transgenic mice for efficient deletion of genes in post-thymic T cells
| Autor: | Ramila Shah, Thomas F. Gajewski, Yuanyuan Zha, Austin Wong, Frederick L. Locke |
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| Rok vydání: | 2007 |
| Předmět: |
Genetically modified mouse
Coxsackie and Adenovirus Receptor-Like Membrane Protein RNA Untranslated Transgene T cell T-Lymphocytes Immunology Genetic Vectors Mice Transgenic Biology medicine.disease_cause Article Viral vector Adenoviridae Transduction (genetics) Mice Transduction Genetic medicine Immunology and Allergy Animals Recombination Genetic PTEN Phosphohydrolase Proteins T lymphocyte Molecular biology In vitro medicine.anatomical_structure Receptors Virus Genetic Engineering Gene Deletion |
| Zdroj: | Journal of immunological methods. 331(1-2) |
| ISSN: | 0022-1759 |
| Popis: | Conditional gene deletion using lineage-specific transgenic expression of Cre has been useful for defining the role of specific gene products in mice in vivo. However, this technology has had limitations for studies of peripheral T cell biology, since the T-lineage promoters commonly used are active early in thymic development. As such, T cell development can be altered by the resulting genetic alterations, thus limiting the interpretation of the data in post-thymic T cell studies. Thus, new strategies are needed to delete targeted genes directly in peripheral T lymphocytes. The availability of transgenic mice expressing the CAR in the T cell compartment enabled testing of Cre-mediated recombination using an adenoviral vector in naive peripheral T cells in vitro, even without cellular activation. Using Rosa26R reporter × CAR transgenic mice, we describe conditions by which Cre-mediated deletion of targeted genes can be achieved with primary T cells in vitro. These cells can also be adoptively transferred into defined recipient mice for study in vivo. We use conditional PTEN-deficient mice as proof of concept to confirm the value of this technique for deleting a negative regulator of T cell activation. This technology should be broadly applicable for studies of T cell-specific gene deletion to gain understanding of function in the post-thymic T cell compartment. |
| Databáze: | OpenAIRE |
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