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Genomic Profiling of Cardiovascular Cell Types During Development
Cardiomyocytes (CM) are the major cardiac cell type. Even so, not much is known regarding the changes that occur during embryonic development as cardiac stem cells differentiate and proliferate to form an adult heart. In order to better understand the changes that occur at different stages of cardiac development, we performed transcriptional profiling of cells during their progression from an immature progenitor state to terminally differentiated cardiomyocytes. Using transgenic mouse models in which cardiac cells can be isolated by a fluorescent reporter protein, we have isolated populations of cardiovascular cells at different stages of development. We generated double transgenic mice by crossing a floxed reporter with cardiac specific-Cre mice. The Cre recombinase was under the transcriptional control of the promoter of Mesp1, Nkx2.5, or α-MHC; markers that are predominantly expressed during the development of the cardiovascular system. The fluorescent-marked cells were then isolated at respective embryonic time points that correlate with the expression of Mesp1, Nkx2.5, or α-MHC. Gene expression analyses were performed using RNA-seq, to identify candidate genes that are potentially important in mediating differentiation and development as well as novel surface markers at each stage. Through the use of MID-REG and deductive reasoning based on gene expression levels we were able to uncover genes that could hold a key role in cardiac cell type development.