Cyclic Amplified Programmable Allosteric DNA Biosensor for Enzyme-Triggered Spatially Controlled Mitochondrial...

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Mitochondrial human apurinic/apyrimidinic endonuclease 1 (APE1) abnormal localization correlates with tumor progression, prognosis, and drug resistance.
• Targeting mitochondrial APE1 for in situ imaging is challenging due to its varied cellular localization (cytoplasm, nucleus, mitochondria).

Purpose of the Study:

• To develop a cyclic amplified programmable allosteric DNA biosensor (C-AP-tFNA) for APE1-triggered, spatially controlled mitochondrial molecular imaging.
• To enable sensitive and specific detection of mitochondrial APE1 in vivo.

Main Methods:

• Designed C-AP-tFNA to interact with cytochrome c, inducing conformational changes for mitochondrial APE1 activation.
• Utilized cyclic amplification and cleavage by mitochondrial APE1 to generate fluorescent signals for imaging.
• Employed an AND-gated mechanism for enhanced specificity.

Main Results:

• C-AP-tFNA demonstrated high specificity and sensitivity for in vivo imaging of mitochondrial APE1 in tumor and inflammatory cells.
• The biosensor successfully monitored neuroblastoma drug resistance in vivo.
• Achieved spatially controlled mitochondrial molecular imaging triggered by APE1.

Conclusions:

• C-AP-tFNA provides a novel and effective approach for sensitive and specific detection of mitochondrial APE1.
• This biosensor is a promising tool for monitoring neuroblastoma drug resistance in vivo.
• The study highlights the potential of programmable DNA biosensors for complex biological imaging challenges.