Simultaneous quantitative assessment of two distinct cell lineages with a nuclear-localized dual genetic reporter. (September 2020) - Tang, Muxue
Abstract: Genetic lineage tracing has been widely used for studying in vivo cell fate plasticity during embryogenesis, tissue homeostasis, and disease development. Recent applications with multiple site-specific recombinases have been used in complex and sophisticated genetic fate mapping studies. However, the previous multicolor reporters for dual recombinases had limitations of precise in situ quantification of cell number, which is mainly due to the intermingling of cells in condensed tissues. Here, we generated a dual recombinase-mediated nuclear-localized GFP and tdTomato reporter line, which enables clear, simultaneous quantification of two distinct cell lineages in vivo . Combining this dual genetic reporter with Tbx18-Cre and Cdh5-Dre lines, which genetically trace epicardial and endothelial cells, respectively, we obtained high-resolution images for the anatomic distribution of the descendants of these two distinct cell lineages in the valve mesenchyme during development, remodeling, and maturation stages. This new dual genetic reporter is expected to facilitate fate tracing of two cell lineages and their objective quantification in vivo . Graphical abstract: Unlabelled Image Highlights: R26-nIR is a dual recombinases-mediated nuclear-localized GFP and tdTomato reporter line R26-nIR enables simultaneous quantification of two distinct cell lineages in vivo R26-nIR obtained high-resolution images for the anatomic distribution of epicardial and endothelial lineages