In vivo gene transfer to the rat retina using herpes simplex virus type 1 (HSV-1)-based amplicon vectors.
The purpose of our study was to evaluate the transduction profiles of herpes simplex virus type 1 (HSV-1)-based amplicon vectors following subretinal injection in the rat. Two amplicon vectors were tested, pHy-CMVGFP and pHy-RPEGFP, both carrying the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) ubiquitous promoter or the RPE65-specific promoter, respectively. For the two amplicon vectors, the GFP reporter gene was efficiently expressed in retinal pigment epithelial (RPE) cells but not in the adjacent photoreceptors. GFP expression was maximum as early as 2 days post-administration but decreased over time to become almost undetectable at 6 weeks postinjection. Super-transduction with a second amplicon vector, pHSVlac, reactivated expression of GFP in approximately 10% of the cells initially transduced at 2 days postinjection of pHy-CMVGFP or pHy-RPEGFP. Reactivation of transgene expression was transient, no GFP signal was detected 8 days after pHSVlac injection. In conclusion, HSV-1 amplicon vectors allow rapid and efficient, but transient, gene transfer in RPE cells following subretinal injection.
Fraefel
C
C
Institute of Virology, University of Zurich, Zurich, Switzerland.
Mendes-Madeira
A
A
Mabon
O
O
Lefebvre
A
A
Le Meur
G
G
Ackermann
M
M
Moullier
P
P
Rolling
F
F
D000818:Animals; D002352:Carrier Proteins; D003587:Cytomegalovirus / Q000235:genetics; D005136:Eye Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics*; D007267:Injections; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000378:metabolism*; D012164:Retinal Diseases / Q000628:therapy; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
D002352:Carrier Proteins; D005136:Eye Proteins; D049452:Green Fluorescent Proteins; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase