Research Report Assessment of CMV, RSV and SYN1 promoters and the woodchuck post-transcriptional regulatory element in adenovirus vectors for transgene expression in cortical neuronal cultures Sherif Boulos a,c, ⁎ , Bruno P. Meloni a,b , Peter G. Arthur c , Christina Bojarski a,b , Neville W. Knuckey a,b a Centre for Neuromuscular and Neurological Disorders, The University of Western Australia, and Australian Neuromuscular Research Institute, Australia b Department of Neurosurgery, Sir Charles Gairdner Hospital, Nedlands, Western Australian, Australia c School of Biomedical and Chemical Sciences, The University of Western Australia, Australia ARTICLE INFO ABSTRACT Article history: Accepted 18 April 2006 Available online 27 June 2006 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. © 2006 Elsevier B.V. All rights reserved. Keywords: Viral vector Promoter Rat cortical neurons Transcriptional enhancer Abbreviations: CMV, cytomegalovirus RSV, rous sarcoma virus hSYN1, human synapsin 1 rSYN1, rat synapsin 1 WPRE, woodchuck post-transcriptional regulatory element Ad, adenovirus EGFP, enhanced green fluorescent protein moi, multiplicity of infection BRAIN RESEARCH 1102 (2006) 27 – 38 ⁎ Corresponding author. Australian Neuromuscular Research Institute, A Block, 4th Floor, QEII Medical Centre, Nedlands, Western Australia, 6009 Australia. Fax: +61 8 9346 3487. E-mail address: sboulos@cyllene.uwa.edu.au (S. Boulos). 0006-8993/$ – see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.brainres.2006.04.089 available at www.sciencedirect.com www.elsevier.com/locate/brainres