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Trends Biotechnol. 2016 Jun;34(6):458-469. doi: 10.1016/j.tibtech.2016.02.009. Epub 2016 Mar 9.

Redefining Signaling Pathways with an Expanding Single-Cell Toolbox.

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Department of Cancer Biology and Center for Cancer Systems Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA. Electronic address:
Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA; Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA. Electronic address:


Genetically identical cells respond heterogeneously to uniform environmental stimuli. Consequently, investigating the signaling networks that control these cell responses using 'average' bulk cell measurements can obscure underlying mechanisms and misses information emerging from cell-to-cell variability. Here we review recent technological advances including live-cell fluorescence imaging-based approaches and microfluidic devices that enable measurements of signaling networks, dynamics, and responses in single cells. We discuss how these single-cell tools have uncovered novel mechanistic insights for canonical signaling pathways that control cell proliferation (ERK), DNA-damage responses (p53), and innate immune and stress responses (NF-κB). Future improvements in throughput and multiplexing, analytical pipelines, and in vivo applicability will all significantly expand the biological information gained from single-cell measurements of signaling pathways.

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