Fluorescence-coded DNA Nanostructure Probe System to Enable Discrimination of Tumor Heterogeneity via a Screening of Dual Intracellular microRNA Signatures in situ

Abstract

Since the delivery kinetics of different cell types are different, the signal from the target cell is greatly affected by the noise signal of the diagnostic system. This is a major obstacle hindering the practical application of intracellular diagnostic systems, such as tumor heterogeneity. To address these issues, here we present a microRNA detection platform using fluorescence-encoded nanostructured DNA-based probes. The nanostructured DNA was designed to include molecular beacons for detecting cytosolic microRNA as well as additional fluorophores. When the intracellular diagnostic system is delivered, fluorescence signals are generated by the molecular beacons, depending on the concentration of the target microRNA. The fluorescence signals are then normalized to the intensity of the additional fluorophore. Through this simple calculation, the concentration of intracellular microRNA can be determined without interference from the diagnosis system itself. And also it enabled discrimination of microRNA expression heterogeneity in five different breast cancer cell lines.