Novel Circular Supercoiling Biosensors for MicroRNA Detection

DESCRIPTION provided by applicant MicroRNA miRNA molecules are single stranded and small nt RNA molecules that act as masters regulators of the genome Research on miRNA has increased dramatically since their discovery just over a decade ago and specific miRNA expression patterns are known to influence most cellular activities including stem cell differentiation and tumor growth However the available techniques to detect miRNA are complex and therefore expensive and time consuming limiting the capacity of researchers and clinicians to validate and use miRNA as disease biomarkers In addition their complicated processes normally introduce sequence bias and make them inaccurate and not reproducible Here we propose the development of a quantitative detection assay based on Circular Supercoiling Biosensors Patent Pending for rapid and multiplex detection of miRNA molecules Circular Supercoiling Biosensors CS Biosensors are a novel kind of biosensor based on DNA supercoiling The new biosensors detect miRNA with single nucleotide specificity without any enzymatic reaction and require only a minute incubation step before electrophoresis detection In this proposal we will optimize CS Biosensors and demonstrate the potential of the biosensors by developing a multiplex miRNA detection panel for tumor analysis and comparing its performance with Taqman RT qPCR The first aim focuses on improving the synthesis and application parameters of the biosensors The second aim focuses on the development and characterization of a method to conduct multiplex miRNA detection Our overall goal is to develop a powerful and flexible miRNA detection kit essential for miRNA research and clinical diagnostics The new kit will be better than available tests for medium and low multiplexing applications targets It will have superior quantitative performance and will be faster simpler to use and less expensive PUBLIC HEALTH RELEVANCE We propose the development of biosensors for the rapid and quantitative analysis of microRNA molecules with high sensitivity and specificity We will demonstrate the new technique by detecting multiple microRNAs in clinical samples using a single reaction