Cre-lox mediated conditional removal of myeloid cell-derived Mif in a mouse model of oral cancer.
MetadataShow full item record
Series/Report no.:Ohio State University. Undergraduate Research Festival. 2020 Autumn.
Oral cancer affects people globally with an estimated 657,000 new cases and 330,000 deaths yearly. Macrophage migration inhibitory factor (Mif) is a gene that encodes for a cytokine involved in regulating cell-mediated immunity, immunoregulation, and inflammation. Previous research has demonstrated that global Mif inhibition results in reduced oral cancer development. This was associated with a decrease in the accumulation of myeloid cell populations in the oral tumor microenvironment. Therefore, our group is interested in elucidating the role of Mif expression by myeloid populations in the oral tumor microenvironment. We previously generated floxed Mif reporter mice (Mif Fl/Fl). Selective breeding between Mif Fl/Fl mice and Cre knock-in mice driven by the Lysozyme promoter (Lyz-Cre) was performed to produce progeny with Mif deficiency in myeloid cells. Genotypic characterization for Mif Fl/Fl and Lyz-Cre was performed by PCR of genomic DNA extracted from mouse ear tissue. Phenotypic characterization of myeloid Mif deficiency was performed through flow cytometric analysis on various myeloid cell populations. Selective breeding of the Mif Fl/Fl mice with the Lyz-Cre mice for three generations yielded myeloid-specific Mif deficient mice. Genotypic PCR characterization of Mif Fl/Fl mice yielded band sizes of 404 bp while wild-type mice yielded a band size of 221 bp. Furthermore, genotypic characterization of Lyz-Cre mice yielded a band size of 700 bp while wild-type mice yielded a band size of 350 bp. Phenotypic flow cytometric analysis of bone marrow-derived macrophages and peritoneal macrophages from myeloid-specific Mif deficient mice revealed an absence of Mif expression in these cells. Furthermore, in these mice, this difference was not observed in lymphoid cells. Our data indicate that using the Cre-LoxP system and selective breeding conditional Mif gene removal in myeloid cells in mice can be achieved to help us gain a better understanding of the role Mif plays in the context of oral cancer and other inflammatory diseases.
Academic Major: Biology
Undergraduate Student Government - Academic Enrichment Grants (AEGs)
Items in Knowledge Bank are protected by copyright, with all rights reserved, unless otherwise indicated.