AbstractHigh precision optical control of neural ensemble activity is essential for understanding brain function and may ultimately revolutionize the treatment of brain disease. Currently only multiphoton holographic optogenetics achieves the requisite spatial resolution for precise neural network control. However, it suffers from critical drawbacks that limit the number of addressable neurons and prevent the development of implantable or wireless devices. Therefore, achieving high-resolution optogenetic control with one-photon activation is essential to catalyze a dramatic leap in spatiotemporally precise multi-site optogenetic technology for research and clinical use. To overcome this challenge, we developed a new light sculpting technique that can synthesize custom illumination patterns in 3D by rapidly projecting structured illumination patterns along with novel computational methods for precise optogenetic control, termed ‘3D-MAP’, for Three-dimensional Multi-site random Access Photostimulation. This technology enables the optogenetic synthesis of complex spatiotemporal sequences of neural activity in the intact brain. As a one-photon photostimulation technology, 3D-MAP can be widely adopted for custom optogenetic applications by the neuroscience research community, and opens the door to scalable, wireless high precision optical brain interfaces.