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Aurora: a fluorescent deoxyribozyme for high-throughput screening.

ABSTRACT: 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.

SUBMITTER: Volek M

PROVIDER: S-EPMC11347150 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature

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Aurora: a fluorescent deoxyribozyme for high-throughput screening.

Volek Martin M Kurfürst Jaroslav J Drexler Matúš M Svoboda Michal M Srb Pavel P Veverka Václav V Curtis Edward A EA

Nucleic acids research 20240801 15

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 ...[more]

PMID: 38860424

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