In vivo transgene expression using an adenoviral tetracycline-regulated system with neuron-specific enolase promoter.
A recombinant adenoviral tetracycline-regulated system with neuron-specific enolase (NSE) promoter was injected stereotaxically into the striatum of rat brains. The efficiency of in vivo transfection was quantified by counting the number of green fluorescent protein (GFP)-positive cells at 3 days, 1 week, and 4 weeks after injection. NeuN immunohistochemistry demonstrated that expression of gammaPKC-GFP was dominant (20-99%) in neuron and expression of gammaPKC-GFP in neuron was significantly higher in pups than adult rats. These results indicate that tetracycline-inhibitable transcription factor (tTA) can drive tetracycline-responsive promoter (TetOp) under the control of NSE promoter, thereby efficiently and selectively expressing gammaPKC-GFP in neurons in vivo.
Bhattacharjee
Abesh Kumar
AK
Department of Neurosurgery, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki Cho, Chuo-Ku, Kobe 650-0017, Japan.
Ueyama
Takehiko
T
Kondoh
Takeshi
T
Hayashi
Shigeto
S
Abouelfetouh
Ayman
A
Sakai
Norio
N
Saito
Naoaki
N
Kohmura
Eiji
E
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000033:anatomy & histology / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D007267:Injections; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism / Q000502:physiology*; D010751:Phosphopyruvate Hydratase / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000502:physiology; D011493:Protein Kinase C / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013238:Stereotaxic Techniques; D013752:Tetracycline / Q000494:pharmacology*; D019076:Transgenes / Q000187:drug effects / Q000235:genetics*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; C087857:protein kinase C gamma; D011493:Protein Kinase C; D010751:Phosphopyruvate Hydratase; D013752:Tetracycline