Studies of Binding Mechanism between Various Types of DNA with Mirna and Its Applications in the Medical Diagnostics and Therapeutics(3/3)

MicroRNAs (miRNAs) are short strands of RNA that act as post-transcriptional regulators. Researchers have found that miRNAs play important role in various disease processes from cancer to cardiovascular disease. When reviewing the potential of miRNAs as biomarkers we need to examine the fundamental problems before can be implemented as routine used. The fundamental problems including the recognizing probe/element design, the transducer, the possible translational issues to the related disease, the easiness and robotics of measurements and etc… To resolve the above-mentioned issues, fundamentally understand the binding/interaction mechanism between the probe (DNA or modified DNA) is pertinent study for miRNA as biomarker.In this study, we will further utilize our research niches built in our lab for the last twenty years on understanding the thermodynamics and kinetics of biomolecular recognition into the binding of miRNA. The probes that we will adapt are regular DNA, Locked nucleic acid (LNA), and peptide nucleic acid (PNA). We will use isothermal titration calorimetry for binding thermodynamics measurements and surface plasmon resonance for binding kinetics. Supported information from circular dichorism, qPCR, and DSC for secondary structure, polymerase extension and Tm (melting temperature) will be determined, respectively. The research results are aim for circulating miRNA profiling and quantitative determination. The binding mechanism study should prove the investigated DNA and modified DNA molecules are recognizing probe for quantitation and profiling of miRNA in the applications of translational medicine.