Source:http://linkedlifedata.com/resource/pubmed/id/17223106
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
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| pubmed:dateCreated |
2007-3-6
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| pubmed:abstractText |
An autoregulated tetracycline-inducible recombinant adeno-associated viral vector (rAAV-pTet(bidi)ON) utilizing the rtTAM2 reverse tetracycline transactivator (rAAV-rtTAM2) was used to conditionally express the human GDNF cDNA. Doxycycline, a tetracycline analog, induced a time- and dose-dependent release of GDNF in vitro in human glioma cells infected with rAAV-rtTAM2 serotype 2 virus. Introducing the Woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) downstream to the rtTAM2 coding sequence, resulted in a more rapid induction and a higher basal expression level. In vivo, 8 weeks after a single injection of the rAAV-rtTAM2-GDNF vector encapsidated into AAV serotype 1 capsids in the rat striatum, the GDNF protein level was 60 pg/mg tissue in doxycycline-treated animals whereas in untreated animals, it was undistinguishable from the endogenous level ( approximately 4 pg/mg tissue). However, a residual GDNF expression in the uninduced animals was evidenced by a sensitive immunohistochemical staining. As compared to rAAV1-rtTAM2-GDNF, the rAAV1-rtTAM2-WPRE-GDNF vector expressed a similar concentration of GDNF in the induced state (with doxycycline) but a basal level (without doxycycline) approximately 2.5-fold higher than the endogenous striatal level. As a proof for biological activity, for both vectors, downregulation of tyrosine hydroxylase was evidenced in dopaminergic terminals of doxycycline-treated but not untreated animals. In conclusion, the rAAV1-rtTAM2 vector which expressed biologically relevant doses of GDNF in the striatum in response to doxycycline with a basal level undistinguishable from the endogenous striatal level, as measured by quantitative ELISA assay, constitutes an interesting tool for local conditional transgenesis.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/Doxycycline,
http://linkedlifedata.com/resource/pubmed/chemical/Glial Cell Line-Derived...,
http://linkedlifedata.com/resource/pubmed/chemical/Green Fluorescent Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Tetracyclines,
http://linkedlifedata.com/resource/pubmed/chemical/Tyrosine 3-Monooxygenase,
http://linkedlifedata.com/resource/pubmed/chemical/enhanced green fluorescent protein
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| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
|
| pubmed:issn |
0014-4886
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| pubmed:author |
pubmed-author:AbeloosLL,
pubmed-author:BlumDD,
pubmed-author:BockstaelOO,
pubmed-author:BrotchiJJ,
pubmed-author:ChtartoAA,
pubmed-author:ESODAE C JEC,
pubmed-author:JasparJ-MJM,
pubmed-author:LehtonenEE,
pubmed-author:LevivierMM,
pubmed-author:MelasCC,
pubmed-author:TenenbaumLL,
pubmed-author:YangXX
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| pubmed:issnType |
Print
|
| pubmed:volume |
204
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| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
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| pubmed:pagination |
387-99
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| pubmed:meshHeading |
pubmed-meshheading:17223106-Animals,
pubmed-meshheading:17223106-Brain,
pubmed-meshheading:17223106-Cell Line, Tumor,
pubmed-meshheading:17223106-Corpus Striatum,
pubmed-meshheading:17223106-DNA, Complementary,
pubmed-meshheading:17223106-Dependovirus,
pubmed-meshheading:17223106-Dose-Response Relationship, Drug,
pubmed-meshheading:17223106-Down-Regulation,
pubmed-meshheading:17223106-Doxycycline,
pubmed-meshheading:17223106-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:17223106-Female,
pubmed-meshheading:17223106-Gene Expression,
pubmed-meshheading:17223106-Gene Transfer Techniques,
pubmed-meshheading:17223106-Genetic Vectors,
pubmed-meshheading:17223106-Glial Cell Line-Derived Neurotrophic Factor,
pubmed-meshheading:17223106-Green Fluorescent Proteins,
pubmed-meshheading:17223106-Humans,
pubmed-meshheading:17223106-Immunohistochemistry,
pubmed-meshheading:17223106-Injections,
pubmed-meshheading:17223106-Rats,
pubmed-meshheading:17223106-Rats, Sprague-Dawley,
pubmed-meshheading:17223106-Tetracyclines,
pubmed-meshheading:17223106-Time Factors,
pubmed-meshheading:17223106-Transduction, Genetic,
pubmed-meshheading:17223106-Transgenes,
pubmed-meshheading:17223106-Tyrosine 3-Monooxygenase
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| pubmed:year |
2007
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| pubmed:articleTitle |
Controlled delivery of glial cell line-derived neurotrophic factor by a single tetracycline-inducible AAV vector.
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| pubmed:affiliation |
Laboratory of Experimental Neurosurgery, Multidisciplinary Research Institute (IRIBHM) Université Libre de Bruxelles, Hôpital Erasme, Belgium.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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