Intracellular microRNA quantification in intact cells: a novel strategy based on reduced graphene oxide-based fluorescen
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D Nanomaterials for Healthcare and Lab-on-a-Chip Devices Prospective Article
Intracellular microRNA quantification in intact cells: a novel strategy based on reduced graphene oxide-based fluorescence quenching Ramasamy Paulmurugan, Cellular Pathway Imaging Laboratory (CPIL), Department of Radiology, Stanford University School of Medicine, 3155 Porter Drive, Suite 2236, Palo Alto, CA 94304, USA Pulickel M. Ajayan, Department of Materials Science and Nanoengineering, Rice University, Houston, TX 77005, USA Dorian Liepmann, Department of Bioengineering, University of California, Berkeley, CA, USA V. Renugopalakrishnan, Boston Children’s Hospital, Harvard Medical School, Boston, MA 02115, USA; Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, USA Address all correspondence to Ramasamy Paulmurugan at [email protected] (Received 26 April 2018; accepted 25 June 2018)
Abstract Nanomaterials have been proposed as key components in biosensing, imaging, and drug delivery since they offer distinctive advantages over conventional approaches. The unique chemical and physical properties of graphene make it possible to functionalize and develop protein transducers, therapeutic delivery vehicles, and microbial diagnostics. In this study, we evaluate reduced graphene oxide as a potential nanomaterial for quantification of microRNAs including their structural differentiation in vitro in solution and inside intact cells. Our results provide evidence for the potential use of graphene nanomaterials as a platform for developing devices that can be used for microRNA quantitation as biomarkers for clinical applications.
Introduction Graphene with its unique electrical and optical properties has been considered important phenomena[1] that can be explored for designing biosensor devices that can specifically and sensitively detect protein, RNA, and DNA biomarkers from various biologic samples. Graphene oxide (GO) and reduced graphene oxide (rGO) are oxidized counterparts of graphene with different electrical and optical properties. In addition to GO’s application in various memory devices and supercapacitors,[2] it is currently used in various biologic applications, which include cellular delivery of drugs, nucleic acids, and proteins, mainly because of high affinity binding of GO for these macromolecules. MicroRNAs are small oligonucleotides of 18–23 base long single-stranded RNAs endogenously expressed in cells and regulate expression levels of various genes.[3,4] MicroRNA biogenesis include several intermediates of single- and double-stranded RNAs, and RNA/DNA hybrids.[5,6] MicroRNA detection and visualization in clinical examples is a powerful approach for a wide range of neoplastic and non-neoplastic diseases. In addition, synthetic microRNA mimics and antisense RNA oligos complementary to endogenous microRNAs are currently used as drugs for treating cellular pathogenesis including cancer and neurodegenerative disorders.[7,8] In addition, microRNAs expressions are dysregulated in cancer and other c
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