Intrinsically Disordered Protein-Inspired Nanovector-Based Coacervates for the Direct Cytosolic Transport of Biomacromolecules

Abstract

In eukaryotic cells, membraneless organelles (MLOs) are formed via liq-uid‒liquid phase separation (LLPS) involving intrinsically disordered proteins(IDPs) and biomacromolecules, enabling biomacromolecule transport withoutvesicles, transporters, or channels. Although MLO-mimetic coacervatesgenerated from synthetic biomaterials can deliver biomacromoleculesinto cells, they lack the conformational adaptability of IDPs and a definedinternalization mechanism, limiting their stability under physiological con-ditions and hindering biomedical translation. Here, IDP-inspired nanovectors(IDP-NVs) are developed with conformational adaptability capable of formingnanocoacervates (NCs) with biomacromolecules for cytosolic delivery. Mixingwith IDP-NVs and cargos results in stable NCs under physiological conditions,and the NCs can directly penetrate cellular membranes through the molecularmotion of IDP-NVs. After the internalization, cytoplasmic glutathionetriggers NC disassembly, releasing biomacromolecules in the cytosol. TheNCs effectively deliver biomacromolecules of diverse sizes, charges, shapes(globular proteins and antibodies), and functions (mRNAs and CRISPR units),demonstrating their versatility and potential for biomedical applications.