Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity
| Autor: | Lauren D. Black, Ayhan Atmanli, François Cherbonneau, Frederik Ernst Deiman, Ibrahim J. Domian, Dongjian Hu, Annebel Marjolein van de Vrugt |
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| Rok vydání: | 2019 |
| Předmět: |
Cell physiology
Myosin light-chain kinase Intravital Microscopy QH301-705.5 Science Cellular differentiation Computational biology Biology General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0302 clinical medicine cellular heterogeneity Gene expression Myosin Fluorescence Resonance Energy Transfer Humans Myocytes Cardiac Biology (General) Induced pluripotent stem cell Gene 030304 developmental biology 0303 health sciences Staining and Labeling General Immunology and Microbiology Gene Expression Profiling General Neuroscience Cell Differentiation Cell Biology General Medicine Tools and Resources cellular differentiation single-cell physiology FRET Medicine lineage diversification Single-Cell Analysis pluripotent stem cells Developmental biology 030217 neurology & neurosurgery Developmental Biology Human |
| Zdroj: | eLife, Vol 8 (2019) eLife |
| ISSN: | 2050-084X |
| DOI: | 10.7554/elife.49599 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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