Targeted gene transfer into developmentally defined cell populations of the primate brain

The primate brain possesses unique physiological and developmental features, yet its systematic investigation has been hampered by a paucity of transgenic germline models and tools. Here, we present a minimally invasive method to introduce transgenes widely across the primate cerebral cortex using ultrasound-guided fetal intracerebroventricular viral injections (FIVIs). FIVI enables efficient and long-lasting transgene expression following intrauterine delivery of recombinant adeno-associated viruses (rAAVs). In the marmoset, we demonstrate that adjusting gestational timing, rAAV serotype, and transcriptional regulatory elements enables selective targeting of defined cell populations, including layer-restricted labeling and Cre-dependent intersectional access. Pilot experiments in rats further demonstrate the potential of FIVIs for prenatal CRISPR-based gene editing and labeling of peripheral somatosensory and retinal pathways. By mimicking key desirable features of germline transgenic models, this efficient and targeted method for gene transfer into the fetal primate brain expands the experimental opportunities for basic and translational neuroscience research across the lifespan.