Genetically Encoded Photoactuators and Photosensors for Characterization and Manipulation of Pluripotent Stem Cells.
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2017
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Abstract
Our knowledge of pluripotent stem cell biology has advanced considerably in the past four decades, but it has yet to deliver on the great promise of regenerative medicine. The slow progress can be mainly attributed to our incomplete understanding of the complex biologic processes regulating the dynamic developmental pathways from pluripotency to fully-differentiated states of functional somatic cells. Much of the difficulty arises from our lack of specific tools to query, or manipulate, the molecular scale circuitry on both single-cell and organismal levels. Fortunately, the last two decades of progress in the field of optogenetics have produced a variety of genetically encoded, light-mediated tools that enable visualization and control of the spatiotemporal regulation of cellular function. The merging of optogenetics and pluripotent stem cell biology could thus be an important step toward realization of the clinical potential of pluripotent stem cells. In this review, we have surveyed available genetically encoded photoactuators and photosensors, a rapidly expanding toolbox, with particular attention to those with utility for studying pluripotent stem cells.
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Pomeroy, Jordan E, Hung X Nguyen, Brenton D Hoffman and Nenad Bursac (2017). Genetically Encoded Photoactuators and Photosensors for Characterization and Manipulation of Pluripotent Stem Cells. Theranostics, 7(14). pp. 3539–3558. 10.7150/thno.20593 Retrieved from https://hdl.handle.net/10161/15568.
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Brenton D. Hoffman
The overall goal of my research program is to utilize an interdisciplinary approach to first advance the basic understanding of mechanotransduction on multiple scales and then use this knowledge to guide the development of new treatments for mechanosensitive diseases. Our work combines principles and techniques from protein engineering, molecular biology, soft matter physics, cell and developmental biology, biomaterials engineering, automated image analysis, and state of the art live cell microscopy. Specifically, we engineer and use biosensors that report the tension across specific proteins in living cells through changes in the color of light they emit. This technology enables dynamic measurements of proteins and sub-cellular structures that are under load. Unlike more traditional techniques that measure the entirety of cellular force output, the ability of these sensors to measure mechanical stress at the molecular level means they are innately compatible with concepts and approaches common in molecular biology and biophysics.
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