Rolling F, Shen W Y, Barnett N L, Tabarias H, Kanagasingam Y, Constable I, Rakoczy P E
Lions Eye Institute, Perth, Western Australia, Australia.
Clin Exp Ophthalmol. 2000 Oct;28(5):382-6. doi: 10.1046/j.1442-9071.2000.00341.x.
Previous studies have demonstrated that adeno-associated virus (AAV) efficiently transduced retinal pigmented epithelial (RPE) cells. The goal of this study was to further evaluate and characterize transgene expression within the RPE cells over time in vivo.
Adeno-associated virus-mediated gene transfer was monitored and quantified by retinal photography following subretinal injection of a recombinant AAV encoding the green fluorescent protein gene (rAAVCMV-gfp) into rat eyes. Retinal function of transduced rat eyes was measured by electroretinography.
The maximum level of transgene expression was reached at 8 weeks postinjection followed by a gradual decrease throughout the experimental period. Interestingly, it was observed that while gfp expression was stable in some RPE cells, gfp fluorescence completely disappeared in other cells over the duration of the experiment. The expression of AAV-mediated gfp in RPE cells did not alter the retinal function for over 1 year
These results confirm the importance of this direct visualization system to study vector transgene expression in vivo and support the use of AAV for diseases treatable by targeting RPE cells.
先前的研究表明，腺相关病毒（AAV）能有效地转导视网膜色素上皮（RPE）细胞。本研究的目的是在体内进一步评估和表征RPE细胞内转基因随时间的表达情况。
将编码绿色荧光蛋白基因的重组腺相关病毒（rAAVCMV - gfp）经视网膜下注射到大鼠眼中后，通过视网膜摄影监测和定量腺相关病毒介导的基因转移。通过视网膜电图测量转导大鼠眼睛的视网膜功能。
注射后8周达到转基因表达的最高水平，随后在整个实验期间逐渐下降。有趣的是，观察到虽然在一些RPE细胞中绿色荧光蛋白（gfp）表达稳定，但在实验过程中其他细胞中的gfp荧光完全消失。RPE细胞中腺相关病毒介导的gfp表达在超过1年的时间里未改变视网膜功能。
这些结果证实了这种直接可视化系统对于研究体内载体转基因表达的重要性，并支持将腺相关病毒用于可通过靶向RPE细胞治疗的疾病。
