Adeno-associated virus vector-mediated triple gene transfer of dopamine synthetic enzymes.
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.


CONCLUSIONS
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.
Fan
D
D
Department of Neurology, Third Hospital, Peking University, Beijing 100083, China. dsfan@sina.com.cn
Shen
Y
Y
Kang
D
D
Nakano
I
I
Ozawa
K
K
D000818:Animals; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics; D004298:Dopamine / Q000096:biosynthesis*; D004798:Enzymes / Q000235:genetics* / Q000378:metabolism; D006136:GTP Cyclohydrolase / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000378:metabolism
D004798:Enzymes; D011993:Recombinant Fusion Proteins; D014446:Tyrosine 3-Monooxygenase; D006136:GTP Cyclohydrolase; D001142:Aromatic-L-Amino-Acid Decarboxylases; D004298:Dopamine