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

Abstract

The primate brain possesses unique physiological and developmental features whose systematic investigation is 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 (FIVI). This technique enables rapid-onset and long-lasting transgene expression following the delivery of recombinant adeno-associated viruses (rAAVs). By adjusting the gestational timing of injections, viral serotypes, and transcriptional regulatory elements, rAAV FIVI allows for systematic targeting of specific cell populations. We demonstrate the versatility of this method through restricted laminar expression in the cortex, Cre-dependent targeting of neurons, 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 opens new avenues for experimental and translational neuroscience across the lifespan.