In situ imaging of proximal biomolecules via Proximity Anchored Modules Assembly

Area of Science:

• Biomolecular imaging
• Molecular diagnostics
• Biotechnology

Background:

• Aberrant proximal biomolecular complexes are crucial disease biomarkers.
• Precise in situ imaging is vital for understanding disease and diagnostics.
• Current imaging methods face limitations in resolution, false positives, and versatility.

Purpose of the Study:

• To introduce Proximity Anchored Modules Assembly (PAMA), a novel imaging strategy.
• To overcome limitations of existing in situ analysis technologies.
• To provide a versatile and multiplexed platform for biomarker imaging.

Main Methods:

• PAMA utilizes a "plug-and-play" architecture decoupling target recognition and signal amplification.
• Programmable DNA tracks synthesized by Primer Exchange Reaction (PER) are employed.
• Polymerase-driven extension is triggered exclusively upon dual-recognition events for proximity-dependent activation.

Main Results:

• PAMA demonstrated precise signal response to homologous and heterologous targets.
• Visualized HER2 receptor dimerization patterns in breast cancer cells.
• Precisely discriminated BCR-ABL^P210 fusion gene isoforms in chronic myeloid leukemia (CML) samples.

Conclusions:

• PAMA offers a rapid (approx. 2.5h), sensitive, and versatile in situ profiling workflow.
• It bridges molecular precision with spatial context for improved diagnostics.
• PAMA establishes a modular framework for precise imaging of diagnostic biomolecular complexes, accelerating discovery and clinical diagnostics.