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rAAV-compatible MiniPromoters for restricted expression in the brain and eye

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Title: rAAV-compatible MiniPromoters for restricted expression in the brain and eye
Authors: De Leeuw, CN
Korecki, AJ
Berry, GE
Hickmott, JW
Lam, SL
Lengyell, TC
Bonaguro, RJ
Borretta, LJ
Chopra, V
Chou, AY
D'Souza, CA
Kaspieva, O
Laprise, S
McInerny, SC
Portales-Casamar, E
Swanson-Newman, MI
Wong, K
Yang, GS
Zhou, M
Jones, SJM
Holt, RA
Asokan, A
Goldowitz, D
Wasserman, WW
Simpson, EM
Item Type: Journal Article
Abstract: Background Small promoters that recapitulate endogenous gene expression patterns are important for basic, preclinical, and now clinical research. Recently, there has been a promising revival of gene therapy for diseases with unmet therapeutic needs. To date, most gene therapies have used viral-based ubiquitous promoters–however, promoters that restrict expression to target cells will minimize off-target side effects, broaden the palette of deliverable therapeutics, and thereby improve safety and efficacy. Here, we take steps towards filling the need for such promoters by developing a high-throughput pipeline that goes from genome-based bioinformatic design to rapid testing in vivo. Methods For much of this work, therapeutically interesting Pleiades MiniPromoters (MiniPs; ~4 kb human DNA regulatory elements), previously tested in knock-in mice, were “cut down” to ~2.5 kb and tested in recombinant adeno-associated virus (rAAV), the virus of choice for gene therapy of the central nervous system. To evaluate our methods, we generated 29 experimental rAAV2/9 viruses carrying 19 different MiniPs, which were injected intravenously into neonatal mice to allow broad unbiased distribution, and characterized in neural tissues by X-gal immunohistochemistry for icre, or immunofluorescent detection of GFP. Results The data showed that 16 of the 19 (84 %) MiniPs recapitulated the expression pattern of their design source. This included expression of: Ple67 in brain raphe nuclei; Ple155 in Purkinje cells of the cerebellum, and retinal bipolar ON cells; Ple261 in endothelial cells of brain blood vessels; and Ple264 in retinal Müller glia. Conclusions Overall, the methodology and MiniPs presented here represent important advances for basic and preclinical research, and may enable a paradigm shift in gene therapy.
Issue Date: 10-May-2016
Date of Acceptance: 30-Apr-2016
URI: http://hdl.handle.net/10044/1/79763
DOI: 10.1186/s13041-016-0232-4
ISSN: 1756-6606
Publisher: BioMed Central
Start Page: 1
End Page: 13
Journal / Book Title: Molecular Brain
Volume: 9
Copyright Statement: © 2016 de Leeuw et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Keywords: Science & Technology
Life Sciences & Biomedicine
Neurosciences
Neurosciences & Neurology
rAAV Gene therapy
Raphe nuclei
Purkinje cells
Retina
Cornea
ADENOASSOCIATED VIRAL VECTORS
OPEN-ACCESS DATABASE
WHOLE-GENOME DATA
GENE-THERAPY
TRANSGENE EXPRESSION
REGULATORY REGIONS
SEROTONIN NEURONS
AAV VECTOR
PROMOTERS
BROWSER
Cornea
Purkinje cells
Raphe nuclei
Retina
rAAV Gene therapy
Animals
Blood-Brain Barrier
Brain
Dependovirus
Dorsal Raphe Nucleus
Eye
Gene Expression
Genetic Vectors
Integrases
Mice, Inbred C57BL
Promoter Regions, Genetic
Recombination, Genetic
Retinal Bipolar Cells
Transduction, Genetic
Eye
Blood-Brain Barrier
Brain
Animals
Mice, Inbred C57BL
Dependovirus
Integrases
Transduction, Genetic
Gene Expression
Recombination, Genetic
Genetic Vectors
Retinal Bipolar Cells
Promoter Regions, Genetic
Dorsal Raphe Nucleus
Science & Technology
Life Sciences & Biomedicine
Neurosciences
Neurosciences & Neurology
rAAV Gene therapy
Raphe nuclei
Purkinje cells
Retina
Cornea
ADENOASSOCIATED VIRAL VECTORS
OPEN-ACCESS DATABASE
WHOLE-GENOME DATA
GENE-THERAPY
TRANSGENE EXPRESSION
REGULATORY REGIONS
SEROTONIN NEURONS
AAV VECTOR
PROMOTERS
BROWSER
Neurology & Neurosurgery
11 Medical and Health Sciences
Publication Status: Published
Article Number: ARTN 52
Online Publication Date: 2016-05-10
Appears in Collections:National Heart and Lung Institute
Faculty of Medicine