Programmable Aggregation of Self-Assembled DNA Constructs.

Area of Science:

• Biochemistry
• Synthetic Biology
• Materials Science

Background:

• Biomolecular aggregates are crucial for cellular biochemical processes.
• Synthetic aggregating systems are vital for mimicking natural dynamic condensates.
• Understanding DNA aggregation is key to developing novel biomaterials.

Purpose of the Study:

• To report ratiometric DNA aggregation of self-assembled DNA constructs using lanthanide salts.
• To demonstrate the reversibility of this aggregation using specific lanthanide-binding ligands.
• To apply this system for evaluating lanthanide-DNA and lanthanide-ligand binding constants.

Main Methods:

• Utilized dynamic light scattering, electrophoretic mobility shift assay, and field emission scanning electron microscopy for aggregate confirmation.
• Employed Forster resonance energy transfer (FRET) pair dyes for tracking binding states.
• Investigated G-quadruplex (GQ) formation using FRET, circular dichroism (CD), and enzyme-linked immunosorbent assay (ELISA).

Main Results:

• Achieved programmed DNA aggregation and reversion using lanthanide salts and specific ligands.
• Successfully evaluated lanthanide-DNA and lanthanide-ligand binding constants.
• Demonstrated a rapid method for lanthanide-induced G-quadruplex formation and stabilization.
• Observed DNase I resistance and distinctive binding to Protoporphyrin dyes and Thioflavin T in lanthanide-DNA constructs.

Conclusions:

• Developed a versatile DNA aggregation system controlled by lanthanides, offering tunable properties.
• Established a new platform for quantifying biomolecular binding interactions.
• Provided a rapid and efficient method for G-quadruplex formation, complementing existing techniques.