Transducing Protease Activity into DNA Output for Developing Smart Bionanosensors

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Biology

Background:

• DNA's information processing is limited to sequence reorganization, lacking input from biological processes.
• Bridging biological events with DNA requires responsive substrates for signal transduction.
• Developing enzyme-to-DNA converters is crucial for expanding DNA's sensing capabilities.

Purpose of the Study:

• To develop and evaluate a two-component sensor for converting protease activity into a DNA-compatible signal.
• To create a DNA-based system capable of discriminating between different proteases.
• To explore the potential of DNA gates for processing multiple biological inputs.

Main Methods:

• Assembled acceptor dye-labeled peptide-DNA onto semiconductor quantum dot (QD) donors as input gates.
• Utilized Förster resonance energy transfer (FRET) to detect protease cleavage of peptide sequences.
• Employed DNA gates for signal transduction and discrimination of protease activity.

Main Results:

• The sensor successfully transduced protease activity into altered FRET signals and DNA output.
• Demonstrated discrimination between trypsin and chymotrypsin based on their cleavage activity.
• Observed FRET sensitization in downstream output gates upon protease interaction.

Conclusions:

• The developed sensor functions as a protease-to-DNA signal converter, enabling biological event detection.
• The DNA-based system exhibits the ability to discriminate between different proteases, enhancing sensing reliability.
• This approach offers a foundation for targeting other enzymes and processing multi-bit biological information.