Characterization of an interferon-induced gene identified through differential display analysis and the generation of knockout mice defective in their ability to produce interferon-induced guanylate-binding proteins

Jill Marinari Carton, Fordham University

Abstract

Differential display analysis was performed on RNA isolated from human Daudi cells that were either untreated or treated with interferon in order to identify interferon-regulated genes. A transcript termed 2-25-A was identified as being up-regulated in interferon-treated Daudi cells. Further studies indicate that this interferon-induced transcript accumulates to elevated levels by four hours after the addition of interferon and the levels remain elevated twenty-four hours post-treatment. The induction of 2-25-A is a direct effect of the interferon treatment and is due to an active increase in the rate of transcription from the 2-25-A gene. In order to study the gene product of the 2-25-A gene, a monoclonal antibody was produced against a portion of the putative 2-25-A protein. This antibody specifically recognizes a protein in cell extracts that is up regulated following interferon treatment. The generation of knockout mice that are defective in their ability to produce interferon-induced guanylate binding proteins was pursued in order to study the role these genes play in interferon's cellular activities. The genomic organization and expression pattern of the murine genes, GBP-1 and PBP, were determined and transgene constructs were designed to specifically target each of these genes. An embryonic stem cell line has been established, for both GBP-1 and PBP that has undergone homologous recombination with the transgene. Increased viral sensitivity has been observed in a cell line mutated at the GBP-1 gene locus.

Subject Area

Molecular biology|Immunology

Recommended Citation

Carton, Jill Marinari, "Characterization of an interferon-induced gene identified through differential display analysis and the generation of knockout mice defective in their ability to produce interferon-induced guanylate-binding proteins" (1998). ETD Collection for Fordham University. AAI9825848.
https://research.library.fordham.edu/dissertations/AAI9825848

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