Aurora: a fluorescent deoxyribozyme for high-throughput screening

0586905 - ÚOCHB 2025 RIV US eng J - Journal Article
Volek, Martin - Kurfürst, Jaroslav - Drexler, Matúš - Svoboda, Michal - Srb, Pavel - Veverka, Václav - Curtis, Edward A.
Aurora: a fluorescent deoxyribozyme for high-throughput screening.
Nucleic Acids Research. (2024). ISSN 0305-1048. E-ISSN 1362-4962
R&D Projects: GA MŠMT(CZ) EH22_008/0004575; GA ČR(CZ) GA24-11210S
Institutional support: RVO:61388963
Keywords : in-vitro selection * DNA aptamer pair * allosteric ribozyme
Impact factor: 14.9, year: 2022
Method of publishing: Open access
https://doi.org/10.1093/nar/gkae467

Fluorescence facilitates the detection, visualization, and tracking of molecules with high sensitivity and specificity. A functional DNA molecule that generates a robust fluorescent signal would offer significant advantages for many applications compared to intrinsically fluorescent proteins, which are expensive and labor intensive to synthesize, and fluorescent RNA aptamers, which are unstable under most conditions. Here, we describe a novel deoxyriboyzme that rapidly and efficiently generates a stable fluorescent product using a readily available coumarin substrate. An engineered version can detect picomolar concentrations of ribonucleases in a simple homogenous assay, and was used to rapidly identify novel inhibitors of the SARS-CoV-2 ribonuclease Nsp15 in a high-throughput screen. Our work adds an important new component to the toolkit of functional DNA parts, and also demonstrates how catalytic DNA motifs can be used to solve real-world problems.
Permanent Link: https://hdl.handle.net/11104/0354277