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Regulating the activity of specific neurons is essentially important in neurocircuit dissection and neuropathy therapy. As a recently developed strategy, nanomaterial‐enabled nongenetic neuromodulations that realize remote physical stimuli have made vast progress and shown great clinical potential. However, minimal invasiveness and high spatiotemporal resolution are still challenging for nongenetic neuromodulation. Herein, a second near‐infrared (NIR‐II)‐light‐induced transcranial nongenetic neurostimulation via bioinspired nanovesicles is reported. The rationally designed vesicles are obtained from vesicle‐membrane‐confined enzymatic reactions. This study demonstrates that the vesicle‐enabled NIR‐II photothermal stimuli can elicit neuronal signaling dynamics with precise spatiotemporal control and thus evoke defined neural circuits in nontransgenic mice. Moreover, the vesicle‐mediated NIR‐II optical stimulation can regulate mouse motor behaviors with minimal invasiveness by eliminating light‐emitting implants. Furthermore, the biological modulation is integrated with photoacoustic brain imaging, realizing navigational, and efficient neuromodulation. Such transcranial and precise NIR‐II optical neuromodulation mediated by bioinspired vesicles shows the potential for the optical‐theranostics of neurological diseases in nontransgenic organisms.

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