Programmable and Chemically Fueled DNA Coacervates by Transient Liquid-Liquid Phase Separation

Area of Science:

• Biochemistry
• Materials Science
• Synthetic Biology

Background:

• Multivalency-driven liquid-liquid phase separation (LLPS) is crucial for biological functions within membrane-less organelles.
• Programmable model systems are needed to understand LLPS and create synthetic reaction environments.

Purpose of the Study:

• To demonstrate a concept for programming LLPS using transient multivalency in ATP-driven sequence-defined functionalized nucleic acid polymers (SfNAPs).
• To create dynamic, multicomponent biomolecular condensate mimics.

Main Methods:

• Utilized an enzymatic reaction network (ERN) of ATP-powered DNA ligation and restriction to form transient SfNAPs.
• Programmed LLPS lifetimes by controlling ATP concentration.
• Engineered distinct molecular recognitions for sorted LLPS and multicomponent coacervates.

Main Results:

• Achieved tunable lifetimes for all-DNA coacervates based on ATP concentration.
• Demonstrated sorted LLPS, creating multicomponent DNA coacervates with programmable recognition.
• Showcased ATP-driven coacervates for trapping colloids and biomolecules, enhancing enzymatic cascade rates.

Conclusions:

• ATP-driven multivalent coacervation is a viable mechanism for dynamic, multicomponent biomolecular condensate mimics.
• This approach offers a valuable tool for autonomous materials design.
• Programmable nucleic acid polymers provide insights into biological LLPS and enable novel synthetic systems.