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.
Tolu
Stefania
S
Unité Neurobiologie Intégrative des Systèmes Cholinergiques, CNRS URA 2182, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.
Avale
M Elena
ME
Nakatani
Hiroko
H
Pons
Stéphanie
S
Parnaudeau
Sébastien
S
Tronche
François
F
Vogt
Angelika
A
Monyer
Hannah
H
Vogel
Roland
R
de Chaumont
Fabrice
F
Olivo-Marin
Jean-Christophe
JC
Changeux
Jean-Pierre
JP
Maskos
Uwe
U
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