Gene transfer into retinal ganglion cells by in vivo electroporation: a new approach

We developed a new in vivo electroporation. method to deliver genes into re tinal ganglion cells (RGCs). Efficiency and degree of tissue damage were ev aluated using green fluorescent protein (GFP) gene and TUNEL. Soon after th e intravitreous injection of the GFP gene, electroporation (five electric p ulses of 99 ms duration each and 12 V/cm. delivered twice 5 min apart) was carried out on the adult rat eyeball with the aid of tweezer-type disc elec trodes attached to corneal (cathode) and scleral (anode) surfaces. GFP expr ession, exhibiting a maximum on day 7, was detectable for up to 21 days. Di I retrograde labeling of RGCs showed that 41.5% of the total ganglion cells in the electroinjected area were GFP-positive. Therefore, this new method may be a useful tool for the delivery of genes into RGCs. (C) 2001 Elsevier Science Ltd. All rights reserved.