The stimuli-responsive CRISPR-Cas12a system for modulating the selective aggregation of cell membrane receptors.

While CRISPR systems exhibit remarkable programmability in the field of nucleic acid editing, their extension to protein engineering faces a fundamental challenge, namely the traditional CRISPR tools lack the design to efficiently convert stimulus signals into the selective clustering of membrane receptors. This study develops a stimulus-responsive membrane-confined CRISPR-Cas12a platform that enhances selective clustering of membrane receptors for functional regulation. Specifically, a membrane-anchored DNA tetrahedral framework (TD-apt) was designed, which leverages vascular endothelial growth factor (VEGF) to activate Cas12a. Compared with unconfined CRISPR-Cas12a, membrane-confined CRISPR-Cas12a exhibits stronger cleavage activity, the interaction between the cellular-mesenchymal epithelial transition factor (c-Met) receptor and transferrin receptor (TfR) on A549 cells was efficiently modulated by nucleic acid assembly. This manipulation selectively inhibited c-Met function through spatial steric hindrance of TfR, modulating cellular behavior. Notably, the system's generality was validated by engineering of c-Met homodimerization for activation. This cascading regulatory paradigm of environmental sensing (VEGF response)-nucleic acid computation (CRISPR-based nucleic acid molecular computation)-protein assembly (receptor topological remodeling) effectively extends CRISPR's application boundaries to the field of non-genetic regulation protein-protein interaction (PPI) and establishes a versatile toolkit for dynamic and precise functional regulation.