Cellular microRNA detection with miRacles: microRNA- activated conditional looping of engineered switches.
Arun Richard ChandrasekaranMolly F MacIsaacParomita DeyOksana LevchenkoLifeng ZhouMadeline AndresBijan K DeyKen HalvorsenPublished in: Science advances (2019)
MicroRNAs are short noncoding regulatory RNAs that are increasingly used as disease biomarkers. Detection of microRNAs can be arduous and expensive and often requires amplification, labeling, or radioactive probes. Here, we report a single-step, nonenzymatic microRNA detection assay using conformationally responsive DNA nanoswitches. Termed miRacles (microRNA-activated conditional looping of engineered switches), our assay has subattomole sensitivity and single-nucleotide specificity using an agarose gel electrophoresis readout. We detect cellular microRNAs from nanogram-scale RNA extracts of differentiating muscle cells and multiplex our detection for several microRNAs from one biological sample. We demonstrate 1-hour detection without expensive equipment or reagents, making this assay a compelling alternative to quantitative polymerase chain reaction and Northern blotting.
Keyphrases
- real time pcr
- loop mediated isothermal amplification
- label free
- high throughput
- blood pressure
- magnetic resonance imaging
- single molecule
- small molecule
- computed tomography
- transcription factor
- magnetic resonance
- drug delivery
- high resolution
- skeletal muscle
- cell proliferation
- single cell
- cell free
- circulating tumor
- fluorescence imaging
- contrast enhanced
- quantum dots