A versatile system for the neuronal subtype specific expression of lentiviral vectors.
Lentiviral expression vectors are powerful tools for gene therapy and long-term gene expression/repression in the mammalian brain. However, no specificity of transduction has been reported so far in the central nervous system. Here we have developed a novel system to achieve a neuronal subtype specific expression in either dopaminergic (DA) or GABAergic neurons. We employed a delivery strategy by which the transgene is not expressed until its activation by Cre recombinase. We successfully tested the system in vitro and then used this novel lentivector, containing loxP sites, in 2 different transgenic mouse lines expressing Cre either in DA or in GABAergic neurons. In both lines the reporter gene was detected exclusively in Cre-positive cells, demonstrating that with this experimental approach we were able to achieve completely specific expression of transgenes delivered by lentiviral vectors. This universal system can be applied to all neural subtypes making use of the growing number of specific Cre driver lines.- Tolu, S., Avale, M. E., Nakatani, H., Pons, S., Parnaudeau, S., Tronche, F., Vogt, A., Monyer, H., Vogel, R., de Chaumont, F., Olivo-Marin, J.-C., Changeux, J.-P., Maskos, U. A versatile system for the neuronal subtype specific expression of lentiviral vectors.
Unité Neurobiologie Intégrative des Systèmes Cholinergiques, CNRS URA 2182, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.
Stefania
S
Tolu
M Elena
ME
Avale
Hiroko
H
Nakatani
Stéphanie
S
Pons
Sébastien
S
Parnaudeau
François
F
Tronche
Angelika
A
Vogt
Hannah
H
Monyer
Roland
R
Vogel
Fabrice
F
de Chaumont
Jean-Christophe
JC
Olivo-Marin
Jean-Pierre
JP
Changeux
Uwe
U
Maskos
D000818:Animals; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D019426:Integrases / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009474:Neurons / Q000378:metabolism; D017557:Ventral Tegmental Area / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D005680:gamma-Aminobutyric Acid; C045073:Cre recombinase; D019426:Integrases; D004298:Dopamine