PT  - JOURNAL ARTICLE
AU  - Liming Wang
AU  - Kern Hast
AU  - Tushar Aggarwal
AU  - Melih Baci
AU  - Jonathan Hong
AU  - Enver Cagri Izgu
TI  - MicroRNA Detection in Biological Media Using a Split Aptamer Platform
AID  - 10.1101/2022.01.17.476684
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.01.17.476684
4099  - http://biorxiv.org/content/early/2022/01/18/2022.01.17.476684.short
4100  - http://biorxiv.org/content/early/2022/01/18/2022.01.17.476684.full
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.