High-level transgene expression in neurons by lentivirus with Tet-Off system.
We developed novel lentiviral vectors by using "Tet-Off system" and succeeded in achieving high-level and neuron-specific gene transduction in vivo. One week after viral injection into the rat neostriatum, the GFP expression was almost completely neuron-specific and about 40 times higher than the expression of a conventional lentiviral vector. High transcriptional activity and neuronal specificity were sustained for up to 8 weeks. Furthermore, neuronal processes of the infected neurons were efficiently visualized by adding a plasma membrane-targeting signal to GFP. These results suggest that the present method is valuable for strong gene transduction and clear visualization of neurons in vivo.
Hioki
Hiroyuki
H
Department of Morphological Brain Science, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Kuramoto
Eriko
E
Konno
Michiteru
M
Kameda
Hiroshi
H
Takahashi
Yasuhiro
Y
Nakano
Takashi
T
Nakamura
Kouichi C
KC
Kaneko
Takeshi
T
D000704:Analysis of Variance; D000818:Animals; D007839:Functional Laterality; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D017072:Neostriatum / Q000166:cytology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*
D009419:Nerve Tissue Proteins; D049452:Green Fluorescent Proteins