Date of Award
Bachelor of Science
Arun Richard Chandrasekaran
MicroRNAs play important roles in gene regulation. Additionally, differential expression of specific microRNAs have been correlated with a wide range of diseases. Sensitive and selective detection of microRNAs is thus important for enabling their use as biomarkers, drugs, or drug targets. Current detection techniques such as northern blotting and quantitative real-time PCR require skilled personnel and expensive equipment to execute complex and time consuming assays. Here we develop and validate a one-step, non-enzymatic microRNA detection assay using DNA nanoswitches programmed to recognize and bind a specific microRNA. Binding induces a loop in the structure, allowing the target microRNA to be unambiguously detected on a standard agarose gel. We demonstrate single nucleotide specificity and sub-attomole sensitivity for synthetic microRNAs in buffer. The utility of the technique is illustrated by biological detection from differentiating muscle cells, where we detect cellular microRNAs from nanogram-scale RNA extracts. Start to finish detection is possible in one hour without expensive equipment or reagents, making this assay a compelling alternative to qPCR and northern blotting.
MacIsaac, Molly F., "One-Step Cellular Micro-RNA Detection with Programmable DNA Nanoswitches" (2018). Biological Sciences. 50.