Assessment of CMV, RSV and SYN1 promoters and the woodchuck post-transcriptional regulatory element in adenovirus vectors for transgene expression in cortical neuronal cultures

Research output: Contribution to journalArticle

35 Citations (Scopus)

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 cytornegalovirus (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 similarbut 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 Bc-X-L overexpression attenuated neuronal death caused by in vitro ischemia and oxidative stress. (c) 2006 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)27-38
JournalBrain Research
Volume1102
Issue number1
DOIs
Publication statusPublished - 2006

Fingerprint

Transcriptional Regulatory Elements
Marmota
Synapsins
Rous sarcoma virus
Transgenes
Adenoviridae
Astrocytes
Neurons
HEK293 Cells
Oxidative Stress
Ischemia
fluorescent protein 583
Proteins

Cite this

@article{38f22828efc943e3be4127c76fa62368,
title = "Assessment of CMV, RSV and SYN1 promoters and the woodchuck post-transcriptional regulatory element in adenovirus vectors for transgene expression in cortical neuronal cultures",
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 cytornegalovirus (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 similarbut 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 Bc-X-L overexpression attenuated neuronal death caused by in vitro ischemia and oxidative stress. (c) 2006 Elsevier B.V. All rights reserved.",
author = "Sherif Boulos and Bruno Meloni and Peter Arthur and Christina Bojarski and Neville Knuckey",
year = "2006",
doi = "10.1016/j.brainres.2006.04.089",
language = "English",
volume = "1102",
pages = "27--38",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Assessment of CMV, RSV and SYN1 promoters and the woodchuck post-transcriptional regulatory element in adenovirus vectors for transgene expression in cortical neuronal cultures

AU - Boulos, Sherif

AU - Meloni, Bruno

AU - Arthur, Peter

AU - Bojarski, Christina

AU - Knuckey, Neville

PY - 2006

Y1 - 2006

N2 - 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 cytornegalovirus (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 similarbut 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 Bc-X-L overexpression attenuated neuronal death caused by in vitro ischemia and oxidative stress. (c) 2006 Elsevier B.V. All rights reserved.

AB - 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 cytornegalovirus (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 similarbut 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 Bc-X-L overexpression attenuated neuronal death caused by in vitro ischemia and oxidative stress. (c) 2006 Elsevier B.V. All rights reserved.

U2 - 10.1016/j.brainres.2006.04.089

DO - 10.1016/j.brainres.2006.04.089

M3 - Article

VL - 1102

SP - 27

EP - 38

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1

ER -