Expression Efficiency of Multiple Il9 Reporter Alleles Is Determined by Cell Lineage

Kharwadkar R; Ulrich BJ; Qayum AA; Koh B; Licona-Limón P; Flavell RA; Kaplan MH (2020). Expression Efficiency of Multiple Il9 Reporter Alleles Is Determined by Cell Lineage 4 (5) :282-291


Generation of allelic gene reporter mice has provided a powerful tool to study gene function in vivo. In conjunction with imaging technologies, reporter mouse models facilitate studies of cell lineage tracing, live cell imaging, and gene expression in the context of diseases. Although there are several advantages to using reporter mice, caution is important to ensure the fidelity of the reporter protein representing the gene of interest. In this study, we compared the efficiency of two Il9 reporter strains Il9citrine and Il9GFP in representing IL-9-producing CD4+ TH9 cells. Although both alleles show high specificity in IL-9–expressing populations, we observed that the Il9GFP allele visualized a much larger proportion of the IL-9–producing cells in culture than the Il9citrine reporter allele. In defining the mechanistic basis for these differences, chromatin immunoprecipitation and chromatin accessibility assay showed that the Il9citrine allele was transcriptionally less active in TH9 cells compared with the wild-type allele. The Il9citrine allele also only captured a fraction of IL-9–expressing bone marrow–derived mast cells. In contrast, the Il9citrine reporter detected Il9 expression in type 2 innate lymphoid cells at a greater percentage than could be identified by IL-9 intracellular cytokine staining. Taken together, our findings demonstrate that the accuracy of IL-9 reporter mouse models may vary with the cell type being examined. These studies demonstrate the importance of choosing appropriate reporter mouse models that are optimal for detecting the cell type of interest as well as the accuracy of conclusions.

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