Dataset Information

Relative Nuclease Resistance of a DNA Aptamer Covalently Conjugated to a Target Protein.

ABSTRACT: A major obstacle to the therapeutic application of an aptamer is its susceptibility to nuclease digestion. Here, we confirmed the acquisition of relative nuclease resistance of a DNA-type thrombin binding aptamer with a warhead (TBA₃) by covalent binding to a target protein in the presence of serum/various nucleases. When the thrombin-inhibitory activity of TBA₃ on thrombin was reversed by the addition of the complementary strand, the aptamer was instantly degraded by the nucleases, showing that the properly folded/bound aptamer conferred the resistance. Covalently binding aptamers possessing both a prolonged drug effect and relative nuclease resistance would be beneficial for in vivo translational applications.

SUBMITTER: Tabuchi Y

PROVIDER: S-EPMC9319527 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

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Relative Nuclease Resistance of a DNA Aptamer Covalently Conjugated to a Target Protein.

Tabuchi Yudai Y Yang Jay J Taki Masumi M

International journal of molecular sciences 20220714 14

A major obstacle to the therapeutic application of an aptamer is its susceptibility to nuclease digestion. Here, we confirmed the acquisition of relative nuclease resistance of a DNA-type thrombin binding aptamer with a warhead (TBA<sub>3</sub>) by covalent binding to a target protein in the presence of serum/various nucleases. When the thrombin-inhibitory activity of TBA<sub>3</sub> on thrombin was reversed by the addition of the complementary strand, the aptamer was instantly degraded by the n ...[more]

PMID: 35887130

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Project description:Electrochemical aptamer-based (E-AB) sensors are a technology capable of real-time monitoring of drug concentrations directly in the body. These sensors achieve their selectivity from surface-attached aptamers, which alter their conformation upon target binding, thereby causing a change in electron transfer kinetics between aptamer-bound redox reporters and the electrode surface. Because, in theory, aptamers can be selected for nearly any target of interest, E-AB sensors have far-reaching potential for diagnostic and biomedical applications. However, a remaining critical weakness in the platform lies in the time-dependent, spontaneous degradation of the bioelectronic interface. This progressive degradation-seen in part as a continuous drop in faradaic current from aptamer-attached redox reporters-limits the in vivo operational life of E-AB sensors to less than 12 h, prohibiting their long-term application for continuous molecular monitoring in humans. In this work, we study the effects of nuclease action on the signaling lifetime of E-AB sensors, to determine whether the progressive signal loss is caused by hydrolysis of DNA aptamers and thus the loss of signaling moieties from the sensor surface. We continuously interrogate sensors deployed in several undiluted biological fluids at 37 °C and inject nuclease to reach physiologically relevant concentrations. By employing both naturally occurring d-DNA and the nuclease-resistant enantiomer l-DNA, we determine that within the current lifespan of state-of-the-art E-AB sensors, nuclease hydrolysis is not the dominant cause of sensor signal loss under the conditions we tested. Instead, signal loss is driven primarily by the loss of monolayer elements-both blocking alkanethiol and aptamer monolayers-from the electrode surface. While use of l-DNA aptamers may extend the E-AB operational life in the long term, the critical issue of passive monolayer loss must be addressed before those effects can be seen.

Dataset Information

Relative Nuclease Resistance of a DNA Aptamer Covalently Conjugated to a Target Protein.

Publications

Relative Nuclease Resistance of a DNA Aptamer Covalently Conjugated to a Target Protein.

Similar Datasets

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