Differential transgene expression profiles in rat brain, using rAAV2/1 vectors with tetracycline-inducible and cytomegalovirus promoters.

Details

Ressource 1Download: BIB_82A1C45B99C0.P001.pdf (2456.34 [Ko])
State: Public
Version: author
Serval ID
serval:BIB_82A1C45B99C0
Type
Article: article from journal or magazin.
Collection
Publications
Title
Differential transgene expression profiles in rat brain, using rAAV2/1 vectors with tetracycline-inducible and cytomegalovirus promoters.
Journal
Human Gene Therapy
Author(s)
Bockstael O., Chtarto A., Wakkinen J., Yang X., Melas C., Levivier M., Brotchi J., Tenenbaum L.
ISSN
1557-7422[electronic], 1043-0342[linking]
Publication state
Published
Issued date
2008
Peer-reviewed
Oui
Volume
19
Number
11
Pages
1293-1305
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Abstract
The biodistribution of transgene expression in the CNS after localized stereotaxic vector delivery is an important issue for the safety of gene therapy for neurological diseases. The cellular specificity of transgene expression from rAAV2/1 vectors (recombinant adeno-associated viral vectors pseudotyped with viral capsids from serotype 1) using the tetracycline-inducible (TetON) expression cassette in comparison with the cytomegalovirus (CMV) promoter was investigated in the rat nigrostriatal pathway. After intrastriatal injection, although green fluorescent protein (GFP) was expressed mainly in neurons with both vectors, the relative proportions of DARPP-32-positive projection neurons and parvalbumin-positive interneurons were, respectively, 13:1 and 2:1 for the CMV and TetON vectors. DARP32-positive neurons projecting to the globus pallidus were strongly GFP positive with both vectors, whereas those projecting to the substantia nigra pars reticulata (SNpr) were efficiently labeled by the CMV vector but poorly by the TetON vector. Numerous GFP-positive cells were evidenced in the subventricular zone with both vectors. However, in the olfactory bulb (OB), GFP-positive neurons were observed with the CMV vector but not the TetON vector. We conclude that the absence of significant amounts of transgene product in distant regions (SN and OB) constitutes a safety advantage of the AAV2/1-TetON vector for striatal gene therapy. Midbrain injections resulted in selective GFP expression in tyrosine hydroxylase-positive neurons by the TetON vector whereas with the CMV vector, GFP-positive cells covered a widespread area of the midbrain. The biodistribution of GFP protein corresponded to that of the transcripts and not of the viral genomes. We conclude that the rAAV2/1-TetON vector constitutes an interesting tool for specific transgene expression in midbrain dopaminergic neurons.
Keywords
Animals, Blotting, Western, Brain/drug effects, Brain/metabolism, Cytomegalovirus/genetics, Dependovirus/genetics, Enzyme-Linked Immunosorbent Assay, Female, Fluorescent Antibody Technique, Gene Expression Regulation/physiology, Gene Therapy, Genetic Vectors, Glial Cell Line-Derived Neurotrophic Factor/genetics, Glial Cell Line-Derived Neurotrophic Factor/metabolism, Immunoenzyme Techniques, Neurons/drug effects, Neurons/metabolism, Promoter Regions, Genetic/genetics, Protein Synthesis Inhibitors/pharmacology, RNA, Messenger/genetics, RNA, Messenger/metabolism, Rats, Rats, Wistar, Reverse Transcriptase Polymerase Chain Reaction, Substantia Nigra/drug effects, Substantia Nigra/metabolism, Tetracycline/pharmacology, Transfection, Transgenes
Pubmed
Web of science
Open Access
Yes
Create date
14/03/2011 13:43
Last modification date
20/08/2019 14:42
Usage data