DNA-Programmed Condensate-Membrane Wetting and Cellular Internalization

Abstract

Deformable condensates offer dynamic interfaces for biomolecule delivery, yet membrane adhesion does not necessarily lead to cellular internalization. The physical transition that determines whether a membrane-bound soft material remains surface-anchored or proceeds through wetting towards productive uptake remains poorly understood, particularly at active living-cell membranes. Here, we engineer sequence-defined DNA condensates through liquid-liquid phase separation (LLPS) and program their interfacial behavior by tuning sticky-end valency and cholesterol organization. These molecular designs precisely regulate condensate fluidity, fusion dynamics and internal organization, generating distinct states of weak contact, persistent anchoring and rapid wetting. Increasing cholesterol-mediated affinity does not enhance uptake. Instead, productive internalization emerges from a balance between membrane adhesion and condensate fluidity and deformability. Native membrane composition further modulates condensate interfacial fate across mammalian cells and plant protoplasts. DNA condensates enrich and deliver CpG ODNs, mRNA ([~]2000 nt) and proteins, while cargo loading experiments reveal that preserving condensate architecture is essential for functional delivery. Our findings identify wetting competence as a design parameter for controlling soft material engagement and cellular entry.