Construction and characterization of adenoviral vector expressing biologically active brain-derived neurotrophic factor.
To deliver brain-derived neurotrophic factor (BDNF) to the central nervous system, we sought to attain adenovirus-mediated transfer and expression of the gene in both in vitro and in vivo experiments. For this purpose, we constructed AxCA-BDNF, a recombinant adenoviral vector containing the BDNF cDNA expression cassette. Reverse transcription polymerase chain reaction analyses of the infected HeLa cells and the transduced mouse brain revealed successful expression of the BDNF gene both in vitro and in vivo. The results of a survival assay of chick dorsal root ganglion cells showed that the produced BDNF was biologically active. We consider, therefore, this newly constructed recombinant adenovirus to be a useful tool to deliver BDNF to degenerating neurons and to be applicable to gene therapy of neurodegenerative diseases and nerve trauma.
Kojima
H
H
Institute of Applied Biochemistry, Gifu, Japan.
Inuzuka
S
S
Miwa
T
T
Furukawa
S
S
Hayashi
K
K
Kanegae
Y
Y
Saito
I
I
Ohishi
N
N
Takamori
M
M
Yagi
K
K
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor; D002470:Cell Survival; D002645:Chickens; D017077:Culture Media, Conditioned; D018076:DNA, Complementary / Q000235:genetics; D005727:Ganglia, Spinal / Q000166:cytology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase; D011994:Recombinant Proteins / Q000096:biosynthesis
D019208:Brain-Derived Neurotrophic Factor; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012194:RNA-Directed DNA Polymerase