Impurity of recombinant adeno-associated virus type 2 affects the transduction characteristics following subretinal injection in the rat.
We recently reported that different purification methods of recombinant adeno-associated virus type 2 (rAAV2) affect the transduction characteristics following subretinal injection. In this study, we examined the roles of contaminant proteins from the HEK-293 cells and helper adenovirus, inactivation of helper adenovirus and cell stress induced by DNA-damaging agents in rAAV-mediated retinal transduction. Our results showed that contaminating factors/proteins resulting from the helper E1 deleted adenovirus are possibly responsible for efficient RPE transduction. Future studies of these factors will undoubtedly lead to development of new therapeutic approaches to PR- and RPE-specific retinal diseases.
Shen
Wei-Yong
WY
Centre for Ophthalmology and Visual Science, The University of Western Australia, 2 Verdun Street, Nedlands, Perth, WA 6009, Australia.
Lai
Yvonne K Y
YK
Lai
Chooi-May
CM
Rakoczy
P Elizabeth
PE
D017498:Adenovirus E1 Proteins / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D002586:Cesium / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D004249:DNA Damage; D000229:Dependovirus / Q000235:genetics*; D005451:Fluorescein Angiography; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins; D006493:Heparin / Q000378:metabolism; D007267:Injections; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D010857:Pigment Epithelium of Eye / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012162:Retinal Degeneration / Q000378:metabolism / Q000628:therapy*; D014161:Transduction, Genetic / Q000379:methods*
D017498:Adenovirus E1 Proteins; D002712:Chlorides; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D002586:Cesium; D006493:Heparin; C028019:cesium chloride