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J Immunol Methods. 2008 Feb 29;331(1-2):94-102. doi: 10.1016/j.jim.2007.11.013. Epub 2007 Dec 26.

Use of Cre-adenovirus and CAR transgenic mice for efficient deletion of genes in post-thymic T cells.

Author information

1
Department of Pathology and Department of Medicine, University of Chicago, Chicago, IL 60637, USA.

Abstract

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 naïve peripheral T cells in vitro, even without cellular activation. Using Rosa26R reporterxCAR 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.

PMID:
18177887
PMCID:
PMC3278801
DOI:
10.1016/j.jim.2007.11.013
[Indexed for MEDLINE]
Free PMC Article

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