研究目的
Investigating the reversible and orthogonal control of a DNAzyme's catalytic function using two distinct photoresponsive molecules for complex DNA computing operations.
研究成果
The study successfully demonstrated the reversible and orthogonal control of a DNAzyme's catalytic function using two distinct photoresponsive molecules, enabling complex DNA computing operations. The system's robustness and potential for applications in DNA nanotechnology and biosensing were highlighted.
研究不足
The system requires optimization for one-pot sequential operation and may benefit from more complex nano-architectures and better light sources for enhanced logic circuit capabilities.
1：Experimental Design and Method Selection
The study involved the incorporation of two distinct photoresponsive molecules (DM-Azo and AAP) into a split horseradish peroxidase mimicking DNAzyme to control its catalytic function reversibly and orthogonally using specific wavelengths of light.
2：Sample Selection and Data Sources
The samples included a split HRP-DNAzyme with a G4 catalytic center consisting of two separated DNA strands, modified with DM-Azo and AAP. Data was collected through UV/Vis spectrometry, fluorescence measurements, and circular dichroism (CD) to monitor the DNAzyme's activity and conformation changes.
3：List of Experimental Equipment and Materials
LED light sources for irradiation at specific wavelengths (350 nm, 365 nm, 450 nm, 590 nm), UV/Vis spectrometer, fluorescence spectrometer, circular dichroism spectrometer.
4：Experimental Procedures and Operational Workflow
The DNAzyme's catalytic activity was controlled by irradiating the samples with specific wavelengths to switch the photoresponsive molecules between their cis and trans conformations. The activity was monitored over multiple switching cycles.
5：Data Analysis Methods
The DNAzyme's activity was quantified by measuring the absorbance at 414 nm to monitor the oxidation of ABTS to ABTS?+. Fluorescence and CD measurements were used to assess hybridization efficiency and G-quadruplex formation, respectively.
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