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Development of photoreceptor‐specific promoters and their utility to investigate EIAV lentiviral vector mediated gene transfer to photoreceptors
Author(s) -
Nicoud Marjorie,
Kong Jian,
Iqball Sharifah,
Kan On,
Naylor Stuart,
Gouras Peter,
Allikmets Rando,
Binley Katie
Publication year - 2007
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1115
Subject(s) - viral vector , gene transfer , promoter , gene , biology , lentivirus , microbiology and biotechnology , virology , genetics , gene expression , human immunodeficiency virus (hiv) , recombinant dna , viral disease
Background We wanted to investigate the ability of recombinant equine infectious anemia virus (EIAV) vectors to transduce photoreceptor cells by developing a series of photoreceptor‐specific promoters that drive strong gene expression in photoreceptor cells. Methods Promoter fragments derived from the rhodopsin ( RHO ), the beta phosphodiesterase ( PDE ) and the retinitis pigmentosa ( RP 1) genes were cloned in combination with an enhancer element, derived from the interphotoreceptor retinoid‐binding protein gene ( IRBP ), into luciferase reporter plasmids. An in vitro transient reporter assay was carried out in the human Y‐79 retinoblastoma cell line. The optimal promoters from this screen were then cloned into the recombinant EIAV vector for evaluation in vivo following subretinal delivery into mice. Results All promoters maintained a photoreceptor‐specific expression profile in vitro and the gene expression was further enhanced in combination with the IRBP enhancer. The use of IRBP‐combined RHO or PDE promoters showed modest but exclusive expression in photoreceptors following subretinal delivery to mice. By contrast an EIAV vector containing the cytomegalovirus (CMV) promoter drove reporter gene expression in both photoreceptors and retinal pigment epithelium. Conclusions It may be possible to use recombinant EIAV vectors containing photoreceptor‐specific promoters to drive therapeutic gene expression to treat a range of retinal degenerative diseases where the photoreceptor cell is the primary disease target. Copyright © 2007 John Wiley & Sons, Ltd.