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腺相关病毒载体在国内猫肾管状上皮细胞中的基因传递。

Gene delivery to renal tubular epithelial cells using adeno-associated virus vector in domestic cats.

机构信息

Sphingolipid Expression Laboratory, Supra-Biomolecular System Group, Frontier Research System, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

出版信息

Res Vet Sci. 2009 Dec;87(3):408-12. doi: 10.1016/j.rvsc.2009.03.016. Epub 2009 Apr 21.

Abstract

Recombinant adeno-associated virus (rAAV) vectors provide excellent gene delivery into the kidney in several mammals. This study evaluated gene delivery into the cat kidney using an rAAV vector. First, infection and reporter gene expression using rAAV vector encoding the enhanced green fluorescent protein gene (rAAV-EGFP) was examined in vitro in epithelial crandell reese feline kidney (CRFK) cells. At 12h after transduction, green fluorescence was detected in cells. Next, the rAAV-EGFP construct was injected into the kidneys of two anesthetized cats via the skin, similar to a renal biopsy. On 3 and 12days after injection, green fluorescence was detected in renal tubules localized near the injected site, but not in glomeruli, blood vessels, or interstitial cells. Finally, the rAAV-EGFP construct was transduced into kidney sections cultured ex vivo. EGFP was expressed in renal tubules between the outer cortex and inner medulla regions. These results demonstrate that rAAV vectors effectively mediate gene delivery into cat renal tubules, and may prove usefulness in gene therapy for cats with renal diseases.

摘要

重组腺相关病毒 (rAAV) 载体在几种哺乳动物中可将基因高效递送至肾脏。本研究利用 rAAV 载体评估了猫肾脏的基因递呈。首先,在体外对转染编码增强型绿色荧光蛋白基因 (rAAV-EGFP) 的 rAAV 载体的猫肾细胞 (CRFK) 进行了感染和报告基因表达检测。转导后 12 小时,细胞中检测到绿色荧光。接下来,通过皮肤将 rAAV-EGFP 构建体注射到两只麻醉猫的肾脏中,类似于肾活检。注射后 3 天和 12 天,在注射部位附近的肾小管中检测到绿色荧光,但在肾小球、血管或间质细胞中未检测到。最后,将 rAAV-EGFP 构建体转导到离体培养的肾脏切片中。EGFP 在肾外皮质和肾髓质区域之间的肾小管中表达。这些结果表明 rAAV 载体可有效介导 cat 肾脏的基因递呈,可能对治疗肾脏疾病的 cat 的基因治疗具有应用价值。

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