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Titolo:
Quantitative analysis of transgene protein, mRNA, and vector DNA followinginjection of an adenoviral vector harboring glial cell line-derived neurotrophic factor into the primate caudate nucleus
Autore:
Kozlowski, DA; Bremer, E; Redmond, DE; George, D; Larson, B; Bohn, MC;
Indirizzi:
Northwestern Univ, Sch Med, Childrens Mem Inst Educ & Res, Chicago, IL 60614 USA Northwestern Univ Chicago IL USA 60614 Educ & Res, Chicago, IL 60614 USA Northwestern Univ, Sch Med, Dept Pediat, Chicago, IL 60614 USA Northwestern Univ Chicago IL USA 60614 Dept Pediat, Chicago, IL 60614 USA Northwestern Univ, Sch Med, Dept Neurosurg, Chicago, IL 60614 USA Northwestern Univ Chicago IL USA 60614 t Neurosurg, Chicago, IL 60614 USA Yale Univ, Sch Med, Dept Psychiat, New Haven, CT 06520 USA Yale Univ New Haven CT USA 06520 , Dept Psychiat, New Haven, CT 06520 USA Yale Univ, Sch Med, Dept Neurosurg, New Haven, CT 06520 USA Yale Univ NewHaven CT USA 06520 Dept Neurosurg, New Haven, CT 06520 USA
Titolo Testata:
MOLECULAR THERAPY
fascicolo: 2, volume: 3, anno: 2001,
pagine: 256 - 261
SICI:
1525-0016(200102)3:2<256:QAOTPM>2.0.ZU;2-U
Fonte:
ISI
Lingua:
ENG
Soggetto:
CENTRAL-NERVOUS-SYSTEM; PARKINSONS-DISEASE; DOPAMINERGIC-NEURONS; SUBSTANTIA-NIGRA; GENE-TRANSFER; GDNF; RAT; EXPRESSION; MODEL; BRAIN;
Keywords:
gene therapy; Parkinson's disease; nigrostriatal system; dopamine neurons; neurotrophic factor; real time RT-PCR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
26
Recensione:
Indirizzi per estratti:
Indirizzo: Bohn, MC Northwestern Univ, Sch Med, Childrens Mem Inst Educ & Res, 2300 Childrens Plaza 209, Chicago, IL 60614 USA Northwestern Univ 2300 Childrens Plaza 209 Chicago IL USA 60614 A
Citazione:
D.A. Kozlowski et al., "Quantitative analysis of transgene protein, mRNA, and vector DNA followinginjection of an adenoviral vector harboring glial cell line-derived neurotrophic factor into the primate caudate nucleus", MOL THER, 3(2), 2001, pp. 256-261

Abstract

Gene therapy for neurodegenerative diseases relies on stable expression ofa vector-mediated transgene in the human central nervous system (CNS). In nonhuman primate CNS, transgene expression has been primarily assessed using descriptive histological methods. Here, we quantified the expression of ahuman glial cell line-derived neurotrophic factor (hGDNF) transgene using an ELISA specific for hGDNF protein and real-time quantitative RT-PCR and PCR for hGDNF mRNA and vector DNA, respectively. Transgene expression was assessed 1 week after injection of an E1-, E3-deleted adenovirus harboring hGDNF into the caudate nucleus of St. Kitts green monkey. We found that 57-147 million and 116-771 million copies of hGDNF mRNA and vector DNA, respectively, were present per 10,000 copies of the beta -actin gene. In the same sites, 40-152 pg of hGDNF protein per milligram of tissue was measured. Comparisons of these measures among monkeys demonstrated variable vector DNA and protein levels, but consistent mRNA levels at one-third of the level of vector DNA. This suggests that local responses to the vector play a role in the level of transgene expression and that high levels of vector DNA do notnecessarily predict a high level of transgene protein. However, the results of this study do show that neuroprotective levels of GDNF transgene expression can be achieved following injection of an adenoviral vector into nonhuman primate caudate. Moreover, these assays provide quantitative methods for evaluating and comparing viral vectors in primate CNS.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/01/20 alle ore 00:32:22