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Assessment of CMV, RSV and SYN1 promoters and the woodchuck post-transcriptional regulatory element in adenovirus vectors for transgene expression in cortical neuronal cultures
- Source :
- Brain research. 1102(1)
- Publication Year :
- 2006
-
Abstract
- In order to investigate protein function in rat primary cortical neuronal cultures, we modified an adenoviral vector expression system and assessed the strength and specificity of the cytomegalovirus (CMV), rous sarcoma virus (RSV), and rat and human synapsin 1 (SYN1) promoters to drive DsRed-X expression. We also incorporated the woodchuck post-transcriptional regulatory element (WPRE) and a CMV promoter-enhanced green fluorescent protein (EGFP) reporter cassette. We observed that the RSV promoter activity was strong in neurons and moderate in astrocytes, while the CMV promoter activity was weak-to-moderate in neurons and very strong in astrocytes. The rat and human SYN1 promoters exhibited similar but weak activity in neurons, despite inclusion of the WPRE. We confirmed that the WPRE enhanced RSV promoter-mediated DsRed-X expression in a time-dependent fashion. Interestingly, we observed very weak SYN1-mediated DsRed-X expression in astrocytes and HEK293 cells suggesting incomplete neuronal-restrictive behavior for this promoter. Finally, using our adenoviral expression system, we demonstrated that RSV promoter-mediated Bcl-X(L) overexpression attenuated neuronal death caused by in vitro ischemia and oxidative stress.
- Subjects :
- Gene Expression Regulation, Viral
Time Factors
Transgene
Blotting, Western
Genetic Vectors
Green Fluorescent Proteins
Cytomegalovirus
Regulatory Sequences, Nucleic Acid
medicine.disease_cause
Viral vector
Gene expression
medicine
Benzene Derivatives
Animals
Hepatitis B Virus, Woodchuck
Humans
Transgenes
Promoter Regions, Genetic
Molecular Biology
Cells, Cultured
Regulation of gene expression
Cerebral Cortex
Neurons
Rous sarcoma virus
biology
Cell Death
General Neuroscience
Promoter
biology.organism_classification
Embryo, Mammalian
Synapsins
Molecular biology
Immunohistochemistry
Rats
Adenoviridae
Avian Sarcoma Viruses
Regulatory sequence
Astrocytes
Neurology (clinical)
Developmental Biology
Subjects
Details
- ISSN :
- 00068993
- Volume :
- 1102
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Brain research
- Accession number :
- edsair.doi.dedup.....e3e8fa91f4fefd3152f2f7f716b1fdb2