Intracameral delivery of AAV to corneal endothelium for expression of secretory proteins

Abstract

AAV9 drives gene expression in a highly selective manner within the corneal endothelium of mice following intracameral inoculation into the anterior chamber of the eye. In principle, this allows genes encoding protein constituents of the secretome (representing up to 20% of the human proteome) to be delivered directly into the aqueous humor. From here the secreted protein moves with the natural flow of the aqueous humor via a pressure gradient and is directed toward the outflow tissues. Such a delivery can be employed to modulate outflow facility and intraocular pressure through interactions at the trabecular meshwork and Schlemm’s canal. We provide a protocol for the delivery of AAV to the corneal endothelium, using a CMV-driven eGFP reporter gene as a marker.