Photodictated Programmed Orthogonal DNA Microdroplets Enable Molecular Communication between Transient Self-Sorting Membraneless Artificial Cells.

Abstract

Bottom-up construction of artificial cells that exhibit dynamic molecular communication, spatial self-sorting, and dissipative signaling remains a central challenge in synthetic biology. In particular, achieving programmable interactions between orthogonal droplet populations is essential for mimicking complex cytomimetic behaviors. Here, we report a photoguided strategy to selectively program the liquid-liquid phase separation (LLPS) of two orthogonal DNA droplet populations assembled from DNA Y-motifs. Furthermore, the two photoguided DNA droplet species undergo transient fusion upon stimulation, followed by dissipation and reformation. During the fusion process, a rapid molecular exchange between the two Y-motifs leads to dynamic molecular communication between the interacting droplets. Upon dissipation of the fused droplets, the exchanged Y-scaffolds spontaneously resegregate into their original droplet populations, revealing an intrinsic self-sorting capability. Building on this dissipative intercommunication mechanism, we demonstrate a multiread and self-rewriting molecular message encryption platform based on neighboring DNA droplets as proof of concept. This work establishes a programmable framework for engineering communicating membraneless artificial cell mimics using dynamic DNA phase-separated systems.