Colorimetric detection of gene transcript by target-induced three-way junction formation

Area of Science:

• Molecular Biology
• Biochemistry
• Analytical Chemistry

Background:

• Alternative splicing generates diverse gene transcripts, crucial for gene expression and biological function.
• Accurate identification of specific transcript variants is essential for precise molecular diagnostics.
• Existing methods may lack the sensitivity or specificity required for complex splicing variations.

Purpose of the Study:

• To develop a simple and highly accurate system for identifying targeted gene transcript variants.
• To leverage G-quadruplex structures for enhanced detection sensitivity.
• To enable precise molecular diagnosis through specific splicing site detection.

Main Methods:

• A three-way junction system was designed using two split G-quadruplex forming probes.
• Probes assemble upon target gene transcript introduction, forming a functional G-quadruplex.
• The G-quadruplex conformation enhances hemin peroxidation for colorimetric detection.

Main Results:

• The system achieved notable resolution for gene splicing site detection.
• The colorimetric assay demonstrated a detection limit of 0.063μM.
• High specificity was achieved, discriminating single-base mismatches around splicing sites.

Conclusions:

• The developed G-quadruplex based system offers a simple and accurate method for gene transcript variant detection.
• The system's sensitivity and specificity support its application in precise molecular diagnostics.
• This approach holds potential for developing simple, equipped devices for gene transcript analysis.