腺相关病毒载体介导的多巴胺合成酶三联基因转移
Adeno-associated virus vector-mediated triple gene transfer of dopamine synthetic enzymes.
作者信息
Fan D, Shen Y, Kang D, Nakano I, Ozawa K
机构信息
Department of Neurology, Third Hospital, Peking University, Beijing 100083, China.
出版信息
Chin Med J (Engl). 2001 Dec;114(12):1276-9.
OBJECTIVE
To explore triple gene transfer of dopamine synthetic enzymes with separate adeno-associated virus (AAV) vectors.
METHODS
The genes for dopamine synthetic enzymes, tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), and GTP cyclohydrolase I (GCH, an enzyme critical for tetrahydrobiopterin synthesis) were contransduced into 293 cells with separate AAV vectors. Expressions of TH, AADC and GCH were detected by Western blot analysis. Intracellular dopamine level was assayed by high-performance liquid chromatography.
RESULTS
TH, AADC and GCH were effectively coexpressed in transduced cells with three separate AAV vectors, AAV-TH, AAV-AADC and AAV-GCH. Furthermore, the coexpression resulted in an effectively spontaneous dopamine production in cotransduced cells.
CONCLUSION
The triple transduction of TH, AADC and GCH genes with separate AAV vectors is effective, which might be important to gene therapy for Parkinson's disease.
目的
探讨使用单独的腺相关病毒（AAV）载体进行多巴胺合成酶的三联基因转移。
方法
将多巴胺合成酶基因，即酪氨酸羟化酶（TH）、芳香族L-氨基酸脱羧酶（AADC）和GTP环化水解酶I（GCH，一种对四氢生物蝶呤合成至关重要的酶），用单独的AAV载体共转导到293细胞中。通过蛋白质印迹分析检测TH、AADC和GCH的表达。通过高效液相色谱法测定细胞内多巴胺水平。
结果
TH、AADC和GCH在转导细胞中与三种单独的AAV载体，即AAV-TH、AAV-AADC和AAV-GCH有效共表达。此外，这种共表达导致共转导细胞中有效自发产生多巴胺。
结论
使用单独的AAV载体对TH、AADC和GCH基因进行三联转导是有效的，这可能对帕金森病的基因治疗具有重要意义。
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