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An Integrated System to Remotely Trigger Intracellular Signal Transduction by Upconversion Nanoparticle-mediated Kinase Photoactivation
Published on: August 30, 2017
A universal CRISPR-Cas14a responsive triple-sensitized upconversion photoelectrochemical sensor
Yu Wang1, Yuan Peng2, Huanying Zhou2
1Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environmental and Operational Medicine, 300050, Tianjin, P.R. China. wangyuyu9210@163.com.
This study introduces a novel CRISPR-Cas14a-based photoelectrochemical (PEC) sensor that combines electrical and fluorescence signals for highly sensitive and stable DNA detection. The developed sensor successfully detected T2 toxin and PTK7 protein at the femtogram level.
Area of Science:
- Biotechnology
- Biosensing
- Molecular Biology
Background:
- CRISPR-Cas14a systems offer high-sensitivity DNA detection and gene editing capabilities.
- Photoelectrochemical (PEC) sensors provide cost-effective and portable analytical tools, but often require improved stability.
- Combining CRISPR-Cas14a with PEC sensing can leverage the strengths of both technologies.
Purpose of the Study:
- To develop a universal CRISPR-Cas14a-responsive ultrasensitive upconversion PEC sensor.
- To enhance the stability and sensitivity of PEC signals using strand displacement amplification (SDA) and near-infrared (NIR) light.
- To demonstrate the sensor's applicability for detecting small molecules and proteins.
Main Methods:
- Development of a modified working electrode (UCNPs-ssDNA-CdS@Au/ITO) as a reporter probe.
- Utilizing the trans-cleavage activity of Cas14a endonuclease.
- Integration of SDA and an NIR light source to improve photoelectric signal stability and sensitivity.
Main Results:
- The developed UCNPs-Cas14a-based PEC sensor achieved a limit of detection in the femtogram (fg) range.
- The sensor demonstrated successful detection of T2 toxin in oats.
- The sensor was also applied for the detection of PTK7 protein in human serum samples.
Conclusions:
- Combining PEC sensing with CRISPR-Cas14a technology creates powerful universal sensors.
- The developed sensor offers ultrasensitive, accurate, and cost-effective detection capabilities.
- This approach holds significant potential for broad applications in detection and diagnostics.
