RT Journal Article
SR Electronic
T1 MicroRNA Detection in Biological Media Using a Split Aptamer Platform
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.01.17.476684
DO 10.1101/2022.01.17.476684
A1 Liming Wang
A1 Kern Hast
A1 Tushar Aggarwal
A1 Melih Baci
A1 Jonathan Hong
A1 Enver Cagri Izgu
YR 2022
UL http://biorxiv.org/content/early/2022/01/18/2022.01.17.476684.abstract
AB Intercellular microRNA (miRNA)-based communication has been implicated in a wide array of functional and dysfunctional biological processes. This has raised attention to the potential use of miRNAs as biomarkers for disease diagnosis and prognosis and produced interest in their detection. Though the list of clinically significant miRNA biomarkers is rapidly expanding, it remains challenging to adapt current tools to investigate new targets in biological environments. Systematic approaches for the rapid development of miRNA biosensors are valuable to reduce this disparity. We describe here a methodology for developing aptamer-based fluorescent biosensors that can specifically detect miRNAs in biological environments, including culture medium from HeLa cells, human serum, and human plasma. This methodology includes the semi-rational design of the hybridization between a pair of split DNA aptamer oligonucleotides and the miRNA target to build a pool of potential sensor designs, and the screening of this pool for designs with high signal-to-background ratio and sequence selectivity. The method uses natural oligonucleotides without chemical modification, and is effective in buffer, 10%, and 30% (v/v) biological media. Following this approach, we developed sensors that detect three miRNA targets (miR-19b, miR-21, and miR-92a) at concentrations as low as 5 nM without amplification and are selective against single-nucleotide mutants. This work expands upon the current design principles of nucleic acid-based biosensors and provides a method to rapidly develop diagnostic tools for novel and niche miRNA targets of interest.Competing Interest StatementThe authors have declared no competing interest.