title: Kinetics of transgene expression in mouse retina following sub-retinal injection of recombinant adeno-associated virus.
abstract: Using confocal microscopy we have examined in detail the temporal and spatial pattern of green fluorescent protein expression following sub-retinal injection of recombinant adeno-associated virus (rAAV) in the mouse and have determined the effect of viral titre on the number and type of cells transduced. Our results suggest that some transgene expression occurs as early as three days after injection, and that transgene expression occurs beyond the area of retinal detachment. Vector titre appears to have a substantial effect on both transduction efficiency and the speed of onset of photoreceptor cell transduction. Our data suggests that we have not yet reached the limits of photoreceptor transduction efficiency using AAV vectors. An increase in titre could still lead to an improved transduction efficiency and faster onset of photoreceptor transduction. We failed to detect transfected cones even in areas where nearly 100% of the rods were transduced, but we found efficient and sustained RPE transduction.
authors.affiliation: Institute of Ophthalmology, University College London, 11-43 Bath Street, EC1V 9EL, London, UK.
authors.forename: G M
authors.initials: GM
authors.lastname: Sarra
authors.forename: C
authors.initials: C
authors.lastname: Stephens
authors.forename: F C
authors.initials: FC
authors.lastname: Schlichtenbrede
authors.forename: J W B
authors.initials: JW
authors.lastname: Bainbridge
authors.forename: A J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: P J
authors.initials: PJ
authors.lastname: Luthert
authors.forename: R R
authors.initials: RR
authors.lastname: Ali
mesh_terms: D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D000229:Dependovirus / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D049452:Green Fluorescent Proteins; D007267:Injections; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008808:Mice, Inbred CBA; D023421:Models, Animal; D010857:Pigment Epithelium of Eye / Q000378:metabolism; D011897:Random Allocation; D012160:Retina / Q000378:metabolism*; D012174:Retinitis Pigmentosa / Q000628:therapy*; D014162:Transfection / Q000379:methods*; D019076:Transgenes
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins