Sequence design-based control of DNA droplets formed from phase separation of DNA nanostructures

TL;DR

This paper presents a novel DNA-based liquid-liquid phase separation system that enables programmable control over droplet formation, dynamics, and interactions through sequence design and enzymatic reactions, advancing applications in synthetic biology.

Contribution

It introduces a new DNA nanostructure system that allows precise control of phase separation behavior and droplet dynamics via sequence design and enzymatic modulation.

Findings

Controlled phase separation temperature through sequence design

Manipulation of droplet fusion, fission, and shape

Protein capture into specific DNA droplets

Abstract

DNA has the potential to realize a controllable liquid-liquid phase separation (LLPS) system, because the design of its base sequences results in programmable interactions. Here, we have developed a novel DNA-based LLPS system which enables us to create 'DNA droplets' and to control their dynamic behaviour by designing sequences of the DNA nanostructure. We were able to change the phase separation temperature required for the formation of DNA droplets by designing the sequences. In addition, the fusion, fission, and formation of Janus-shaped droplets were controlled by sequence design and enzymatic reactions. Furthermore, modifications of proteins with sequence-designed DNAs allowed for their capture into specific droplets. Overall, our results provide a new platform for designing the phase behaviour of macromolecular structures, and paves the way for new applications of sequence-designed DNA in the creation of cell-mimicries, synthetic membraneless organelles, and artificial molecular systems.