Subzymes: Regulating DNAzymes for point of care nucleic acid sensing
| Autor: | Radhika Ramadas, Nicole J. Hasick, Alison V. Todd |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Chemistry
Oligonucleotide Metals and Alloys Deoxyribozyme Substrate (chemistry) 02 engineering and technology 010402 general chemistry 021001 nanoscience & nanotechnology Condensed Matter Physics Cleavage (embryo) 01 natural sciences Combinatorial chemistry 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials Catalysis Materials Chemistry Nucleic acid Electrical and Electronic Engineering 0210 nano-technology Instrumentation Signal amplification Point of care |
| Zdroj: | Sensors and Actuators B: Chemical. 297:126704 |
| ISSN: | 0925-4005 |
| Popis: | Rapid, sensitive and affordable nucleic acid sensors that can operate in a point of care (POC) setting are highly desired; however, their availability and implementation remain limited. We report the development of a strategy for building sensors which use novel catalytic nucleic acid structures, herein referred to as ‘Subzymes’. We examine the effectiveness of Subzymes to mediate signal amplification in a universal manner allowing faster rates of detection. We created Subzymes by manufacturing composite oligonucleotides that contain a catalytic nucleic acid component and a substrate component. We demonstrate that the activity of some catalytic DNAzyme components can be inhibited by attaching the Subzymes to micro-particles. Subsequent cleavage of a Subzyme’s internal substrate results in the release and activation of the surface-bound DNAzyme, thus providing a mechanism to control the catalytic activity of the DNAzyme. We demonstrate that released DNAzymes are capable of cleaving fluorescent-labelled reporter substrates to generate a signal, thus confirming the restoration of their catalytic activity. The addition of Subzymes to reactions where the detection of a target is achieved by multi-component nucleic acid enzymes, known as PlexZymes, showed improved target sensitivities in a shorter amount of time (10 pM of target detected in under 60 min), demonstrating rapid detection of nucleic acid targets without the use of protein-enzymes. Subzymes are constructed from low-cost materials and operate under isothermal reaction conditions which are advantageous for on-site diagnostic testing. Thus, Subzymes offer a universal, rapid and affordable tool for nucleic acid sensing, providing new avenues for POC testing. |
| Databáze: | OpenAIRE |
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