Atmanli, Ayhan;
Hu, Dongjian;
Deiman, Frederik Ernst;
van de Vrugt, Annebel Marjolein;
Cherbonneau, François;
Black, Lauren Deems;
Domian, Ibrahim John
Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity
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Medientyp:
E-Artikel
Titel:
Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity
Beteiligte:
Atmanli, Ayhan;
Hu, Dongjian;
Deiman, Frederik Ernst;
van de Vrugt, Annebel Marjolein;
Cherbonneau, François;
Black, Lauren Deems;
Domian, Ibrahim John
Beschreibung:
<jats:p>A fundamental goal in the biological sciences is to determine how individual cells with varied gene expression profiles and diverse functional characteristics contribute to development, physiology, and disease. Here, we report a novel strategy to assess gene expression and cell physiology in single living cells. Our approach utilizes fluorescently labeled mRNA-specific anti-sense RNA probes and dsRNA-binding protein to identify the expression of specific genes in real-time at single-cell resolution via FRET. We use this technology to identify distinct myocardial subpopulations expressing the structural proteins myosin heavy chain α and myosin light chain 2a in real-time during early differentiation of human pluripotent stem cells. We combine this live-cell gene expression analysis with detailed physiologic phenotyping to capture the functional evolution of these early myocardial subpopulations during lineage specification and diversification. This live-cell mRNA imaging approach will have wide ranging application wherever heterogeneity plays an important biological role.</jats:p>