Abstract
Environmental pH has a determining role in the structure of biomolecules, thus playing an important role in regulating cellular activities. Eukaryotic cells must, therefore, strive to stringently regulate pH in various intracellular organelles so as to confer normal functioning at the level of whole organism. Several pH-sensitive probes have been reported, each of which can be used to map the pH dependence of an in vivo process. However, these probes suffer from some inherent drawbacks. Here we demonstrate the utility of an externally introduced, pH-triggered DNA nanomachine inside the multicellular eukaryote Caenorhabditis elegans. The nanomachine uses FRET to effectively map spatiotemporal pH changes associated with endocytosis in coelomocytes of wild type as well as mutant worms, in a variety of genetic backgrounds. It shows highest dynamic range in the pH regime 5.3–6.6 and has a half-life of ∼8 h, thus positioning it well to interrogate a variety of pH-correlated biological phenomena in vivo.
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Acknowledgments
We thank Sandhya P. Koushika for inputs on experiments, Souvik Modi for technical input, Central Imaging Facility at NCBS and the Caenorhabditis Genetics Center (funded by NIH-NCRR) for nematode strains, and DBT and the Nanoscience and Technology Initiative of DST for funding. S.S. acknowledges the CSIR, and Y.K. acknowledges the Innovative Young Biotechnologist Award and Wellcome Trust–DBT India Alliance for fellowships.
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Surana, S., Krishnan, Y. (2013). A Method to Map Spatiotemporal pH Changes in a Multicellular Living Organism Using a DNA Nanosensor. In: Weissig, V., Elbayoumi, T., Olsen, M. (eds) Cellular and Subcellular Nanotechnology. Methods in Molecular Biology, vol 991. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-336-7_2
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DOI: https://doi.org/10.1007/978-1-62703-336-7_2
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