title: Adenovirus vector-mediated in vivo gene transfer into adult murine retina.
abstract: OBJECTIVE
  To determine whether a reporter gene can be introduced into the adult mammalian retina in vivo through means of a recombinant replication-deficient adenovirus.
  
  
  METHODS
  A dilution series of purified Ad.CMVlacZ ranging from 10(5) to 10(11) pfu/ml was prepared and microinjected into the subretinal space of adult CD-1 mice. This virus contained the cytomegalovirus (CMV)-promoted Escherichia coli reporter gene, lacZ. LacZ expression was assessed in enucleated eyes from 0 to 95 days after injection by beta-galactosidase (beta-Gal) assay.
  
  
  RESULTS
  The efficiency of transfection increased as a function of concentration of recombinant virus injected. Eyes injected with greater than 10(7) pfu of Ad.CMVlacZ demonstrated beta-Gal activity lasting at least 95 days. LacZ expression was apparent only in those cells directly exposed to the adenovirus. LacZ expression was observed in the retinal pigment epithelium (RPE) at high efficiency at 48 hours after exposure. By 2 weeks after injection of > 10(7) pfu, lacZ was also expressed in photoreceptors, but at lower density.
  
  
  CONCLUSIONS
  These results demonstrate that high efficiency stable transfer of functional genes can be achieved in vivo in post-mitotic mammalian retina using recombinant adenoviral vectors. Adenovirus vectors appear to be a promising means for delivering therapeutic genes in vivo to the mammalian neural retina and particularly to the RPE.
authors.affiliation: Scheie Eye Institute, F. M. Kirby Center for Molecular Ophthalmology, Philadelphia, PA 19103.
authors.forename: J
authors.initials: J
authors.lastname: Bennett
authors.forename: J
authors.initials: J
authors.lastname: Wilson
authors.forename: D
authors.initials: D
authors.lastname: Sun
authors.forename: B
authors.initials: B
authors.lastname: Forbes
authors.forename: A
authors.initials: A
authors.lastname: Maguire
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D003673:Defective Viruses / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006651:Histocytochemistry; D007763:Lac Operon; D051379:Mice; D010786:Photoreceptor Cells / Q000201:enzymology; D010857:Pigment Epithelium of Eye / Q000201:enzymology; D012160:Retina / Q000201:enzymology; D012163:Retinal Detachment / Q000201:enzymology; D013997:Time Factors