title: Optimizing promoters for recombinant adeno-associated virus-mediated gene expression in the peripheral and central nervous system using self-complementary vectors.
abstract: With the increased use of small self-complementary adeno-associated viral (AAV) vectors, the design of compact promoters becomes critical for packaging and expressing larger transgenes under ubiquitous or cell-specific control. In a comparative study of commonly used 800-bp cytomegalovirus (CMV) and chicken β-actin (CBA) promoters, we report significant differences in the patterns of cell-specific gene expression in the central and peripheral nervous systems. The CMV promoter provides high initial neural expression that diminishes over time. The CBA promoter displayed mostly ubiquitous and high neural expression, but substantially lower expression in motor neurons (MNs). We report the creation of a novel hybrid form of the CBA promoter (CBh) that provides robust long-term expression in all cells observed with CMV or CBA, including MNs. To develop a short neuronal promoter to package larger transgenes into AAV vectors, we also found that a 229-bp fragment of the mouse methyl-CpG-binding protein-2 (MeCP2) promoter was able to drive neuron-specific expression within the CNS. Thus the 800-bp CBh promoter provides strong, long-term, and ubiquitous CNS expression whereas the MeCP2 promoter allows an extra 570-bp packaging capacity, with low and mostly neuronal expression within the CNS, similar to the MeCP2 transcription factor.
authors.affiliation: Gene Therapy Center, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Gray
authors.forename: Stacey B
authors.initials: SB
authors.lastname: Foti
authors.forename: Joel W
authors.initials: JW
authors.lastname: Schwartz
authors.forename: Lavanya
authors.initials: L
authors.lastname: Bachaboina
authors.forename: Bonnie
authors.initials: B
authors.lastname: Taylor-Blake
authors.forename: Jennifer
authors.initials: J
authors.lastname: Coleman
authors.forename: Michael D
authors.initials: MD
authors.lastname: Ehlers
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Zylka
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: McCown
authors.forename: R Jude
authors.initials: RJ
authors.lastname: Samulski
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D002490:Central Nervous System / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005260:Female; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000378:metabolism; D009928:Organ Specificity / Q000235:genetics; D017933:Peripheral Nervous System / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic
chemical_list: D000199:Actins; D051783:Methyl-CpG-Binding Protein 2