Phase-separating enzyme-responsive condensates for inhalable delivery of biomacromolecules

Inhaled delivery of biomacromolecules shows extensive promise for treating a large variety of diseases, especially pulmonary and respiratory diseases. Although nanoparticle-based vehicles enhance intracellular uptake efficiency, it still remains challenging for biomacromolecules inhaled delivery to overcome cytotoxicity, cargo leakage and off-target effect. Here, we present an enzyme-responsive condensates based highly biocompatible and controllable platform for inhaled delivery of biomacromolecules. The condensates that can be modularly designed for rapid recruitment and enrichment of biomolecules, including Cas9 ribonucleoprotein (RNP) and mRNA, were internalized by cells via lysosomal-independent endocytosis, and subsequently released payloads triggered with endogenous physicochemical stimulus for controlled regulation of genes and proteins expression. Additionally, the micrometer sized condensates with highly structural and biological stability during lyophilization and nebulization enable inhalable delivery of biomacromolecules to lung. This condensate-based vehicle provides a promising platform for biomacromolecules inhaled delivery and controlled release, targeted cell regulation, and precise therapy.