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Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity.


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 ? 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.

SUBMITTER: Atmanli A 

PROVIDER: S-EPMC6861004 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity.

Atmanli Ayhan A   Hu Dongjian D   Deiman Frederik Ernst FE   van de Vrugt Annebel Marjolein AM   Cherbonneau François F   Black Lauren Deems LD   Domian Ibrahim John IJ  

eLife 20191008


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 resolut  ...[more]

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