Precise Control of Intracellular Trafficking and Receptor‐Mediated Endocytosis in Living Cells and Behaving Animals

Intracellular trafficking, an extremely complex network, dynamically orchestrates nearly all cellular activities. A versatile method that enables the manipulation of target transport pathways with high spatiotemporal accuracy in vitro and in vivo is required to study how this network coordinates its functions. Here, a new method called RIVET (Rapid Immobilization of target Vesicles on Engaged Tracks) is presented. Utilizing inducible dimerization between target vesicles and selective cytoskeletons, RIVET can spatiotemporally halt numerous intracellular trafficking pathways within seconds in a reversible manner. Its highly specific perturbations allow for the real‐time dissection of the dynamic relationships among different trafficking pathways. Moreover, RIVET is capable of inhibiting receptor‐mediated endocytosis. This versatile system can be applied from the cellular level to whole organisms. RIVET opens up new avenues for studying intracellular trafficking under various physiological and pathological conditions and offers potential strategies for treating trafficking‐related disorders. The RIVET (Rapid Immobilization of Target Vesicles on Engaged Tracks) system enables spatiotemporal perturbation of specific intracellular trafficking pathways and receptor‐mediated endocytosis in living cells and organisms. This is accomplished through chemically or light‐inducible dimerization between target vesicles, granules, or receptors and their associated railways, such as microtubules and actin filaments. This powerful tool allows for the dissection of dynamic intracellular trafficking processes and holds potential therapeutic applications, such as blocking viral entry.