Abstract
Optogenetics is an emerging technology for the manipulation and control of excitable tissues, such as the brain and heart. As this technique requires the genetic modification of cells in order to inscribe light sensitivity, for cardiac applications, here we describe the process through which neonatal rat ventricular myocytes are virally infected in vitro with channelrhodopsin-2 (ChR2). We also describe in detail the procedure for quantitatively determining the optimal viral dosage, including instructions for patterning gene expression in multicellular cardiomyocyte preparations (cardiac syncytia) to simulate potential in vivo transgene distributions. Finally, we address optical actuation of ChR2-transduced cells and means to measure their functional response to light.
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Acknowledgements
This work was supported by National Heart, Lung, and Blood Institute Grant R01-HL-111649 (E.E.), an Institutional National Service Research Award T32-DK07521 (C.M.A.), and partially a NYSTEM grant C026716 to the Stony Brook Stem Cell Center.
We would also like to thank Varsha Sitaraman, PhD; Jinzhu Yu, BS; and Kay Chen, BS, for their contributions.
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Ambrosi, C.M., Entcheva, E. (2014). Optogenetic Control of Cardiomyocytes via Viral Delivery. In: Radisic, M., Black III, L. (eds) Cardiac Tissue Engineering. Methods in Molecular Biology, vol 1181. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1047-2_19
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DOI: https://doi.org/10.1007/978-1-4939-1047-2_19
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