Gene delivery to renal tubular epithelial cells using adeno-associated virus vector in domestic cats.
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.
Miyazaki
Masao
M
Sphingolipid Expression Laboratory, Supra-Biomolecular System Group, Frontier Research System, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. mmiyazaki@tokai-u.jp
Yamashita
Tetsuro
T
Miyazaki
Tamako
T
Taira
Hideharu
H
Suzuki
Akemi
A
D000256:Adenoviridae; D000818:Animals; D002415:Cats; D002460:Cell Line; D015316:Genetic Therapy / Q000662:veterinary*; D049452:Green Fluorescent Proteins; D007668:Kidney / Q000166:cytology / Q000502:physiology*; D008297:Male; D046509:Tissue Culture Techniques
D049452:Green Fluorescent Proteins