Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity
- Massachusetts General Hospital, United States
- Tufts University, United States
Abstract
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.
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All data generated or analysed during this study are included in the manuscript and supporting files.
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Author details
Funding
American Heart Association (Predoctoral Fellowship)
- Ayhan Atmanli
National Heart, Lung, and Blood Institute (Progenitor Cell Biology Consortium (PCBC) Jump Start Award)
- Ayhan Atmanli
National Heart, Lung, and Blood Institute (U01HL100408-01)
- Ibrahim Domian
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2019, Atmanli et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity
eLife 8:e49599.
https://doi.org/10.7554/eLife.49599
