Intraocular injection of adeno-associated viral (AAV) vectors has been an evident
route for delivering
gene drugs into the
retina. Currently, the vectors need to be injected into the subretinal spacein order to provide
gene delivery to cones. In this approach,
gene delivery is limited to cells that contact the local "bleb" of injected fluid. Furthermore,
retinal detachment that occurs during subretinal injections is a concern in eyes with
retinal degeneration. Here, the inventors establish several new vector-
promoter combinations to overcome the limitations associated with AAV-mediated cone transduction in the fovea with supporting studies in mouse models, human
induced pluripotent stem cell-derived organoids, post-mortem human
retinal explants and living macaques. They show that an AAV9variant provides efficient foveal cone transduction when injected into the subretinal space several millimeters away from the fovea, without detaching this delicate region. The delivery modality relies on a cone-specific
promoter and result in high-level
transgene expression compatible with optogenetic vision restoration. Accordingly, the present invention relates to method of expressing a
polynucleotide of interest in the cone photoreceptors of a subject comprising subretinal delivery of a therapeutically effective amount of a recombinant AAV9-derived vector comprising a VP1
capsid protein asset forth in SEQ ID NO: 11 and the
polynucleotide of interest under the control of the pR1.7
promoter as set forth in SEQ ID NO: 12.