Nucleic Acid Detection by a Target-Assisted Proximity Proteolysis Reaction

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Nucleic acid analysis is crucial for disease diagnosis and biological research.
• Current methods for nucleic acid detection often lack speed, simplicity, or portability, especially for remote or low-resource settings.

Purpose of the Study:

• To develop a rapid, simple, and sensitive method for detecting specific DNA and RNA sequences.
• To establish a proximity-enhanced proteolysis reaction as a novel platform for nucleic acid detection.

Main Methods:

• Site-specific modification of a protease and its zymogen with oligonucleotides.
• Utilizing the proximity-enhanced proteolysis reaction triggered by target nucleic acid binding.
• Detection of nucleic acids via an absorbance signal.

Main Results:

• Detection of target DNA and RNA in under 1 hour at sub-nanomolar concentrations.
• The assay demonstrated resistance to interference from biological matrices.
• Sensitivity was enhanced when coupled with isothermal nucleic acid amplification.

Conclusions:

• The proximity proteolysis reaction offers a feasible new platform technology for specific nucleic acid sequence detection.
• This method holds promise for applications in resource-limited environments and point-of-care diagnostics.