Bicycronic construct for optogenetic prosthesis of ganglion cell receptive field of degenerated retina

For the purpose of optogenetic prosthetics of the receptive field of the retinal ganglion cell, we have created a bicistronic genetic construct that carries genes of excitatory (channelorhodopsin2) and inhibitory (anionic channelorhodopsin) rhodopsins. A distinctive feature of this construct is the combination of two genes into one construct with the mutant IRES inserted between them, which ensures precise ratio of the expression levels of the first and second gene in each transfected cell. It was found that the illumination of the central part of transfected neuron with light with a wavelength of 470 nm causes the generation of action potentials in the cell. At the same time, light stimulation of the periphery of the neuron causes cessation of the generation of action potentials. Thus, we were able to simulate the ON-OFF interaction of the receptive field of the retinal ganglion cell using optogenetic methods. Theoretically, this construction can be used for optogenetic prosthetics of degenerative retina in case of its delivery to ganglion cells using lentiviral vectors.