Phase-Transformable DNA Frameworks for Synthetic Condensates with Valency-Controlled Subcellular Sorting

Area of Science:

• Synthetic biology
• Biophysics
• Nanotechnology

Background:

• Controlling intracellular phase separation is key for engineering artificial membraneless organelles (MLOs).
• Precise control of DNA-based condensates within cellular environments is challenging.

Purpose of the Study:

• To develop a phase-transforming DNA framework for in situ formation of functional condensates.
• To investigate the role of binding valencies in controlling condensation order and intracellular fate.
• To demonstrate the application of programmable DNA condensates as artificial MLOs.

Main Methods:

• Utilized coarse-grained simulations and experimental approaches.
• Employed valency-controlled subcellular sorting of tetrahedral DNA nanostructures.
• Analyzed intracellular fates of synthetic DNA (synDNA) condensates using high- and low-order structures.

Main Results:

• Demonstrated that condensation order is dictated by the binding valencies of building blocks.
• Revealed distinct intracellular fates: high-order condensates evade the endolysosomal pathway, while low-order ones are lysosome-trapped.
• Showcased spatially controlled DNA condensates for targeted membrane protein degradation and prolonged therapeutic payload residence.

Conclusions:

• Established a modular design principle for creating programmable DNA condensates in cellulo.
• Highlighted the significant interactions between synthetic constructs and cellular organelles.
• Opened avenues for advanced synthetic biology applications using DNA-based condensates.