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Volume 11, Issue 2, Pages 275-283 (February 2005)
Biodistribution of rAAV Vectors Following Intraocular Administration: Evidence for the Presence and Persistence of Vector DNA in the Optic Nerve and in the Brain Nathalie Provost, Guylène Le Meur, Michel Weber, Alexandra Mendes-Madeira, Guillaume Podevin, Yan Cherel, Marie-Anne Colle, Jack-Yves Deschamps, Philippe Moullier, Fabienne Rolling Molecular Therapy Volume 11, Issue 2, Pages (February 2005) DOI: /j.ymthe Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
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Fig. 1 The GFP expression in rAAVCMV.gfp-injected animals. (A, B, C, and D) Live fluorescence retinal imaging and (E and F) neuroretina flat-mount examined under inverted fluorescence microscope. (A) A rAAV-2/2.CMV.gfp subretinally injected rat (SR1) at 1 month p.i., (B) a rAAV-2/2.CMV.gfp intravitreally injected rat (IV1) at 1 month p.i., (C) a rAAV-2/4.CMV.gfp subretinally injected primate (PSR1) at 2 months p.i., (D) a rAAV-2/5.CMV.gfp subretinally injected dog (DSR3) at 16 months p.i., and (E and F) a rAAV-2/2.CMV.gfp intravitreally injected dog (DIV1) at 4 months p.i. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
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Fig. 2 Vector dissemination to various organs and tissues following subretinal or intravitreal delivery of rAAV-2/2.CMV.gfp in rats. (A) Subretinally injected rat SR5. (B) Intravitreally injected rat IV6. Genomic DNA was extracted from each tissue and analyzed by PCR. PCR was carried out using primers that amplified a 424-bp region within the Gfp sequence. Reaction products were separated on an agarose gel (top) and then transferred to a nylon membrane and hybridized to a Gfp probe (bottom). For IV6 (B), a longer exposure is shown. As a control, each sample was also used for the amplification of a 350-bp cytochrome b DNA fragment. Also shown are the DNA marker (M), a positive control on 25 pg of vector plasmid (+), and a negative control on water (−). Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
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Fig. 3 Vector dissemination to various organs and tissues following subretinal delivery of rAAV-2/4.CMV.gfp in a nonhuman primate (PSR1). The PCR analysis was carried out as described. The DNA marker (M), positive control on 25 pg of vector plasmid (+), and negative control on water (−) are included. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
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Fig. 4 Vector dissemination to various organs and tissues following intravitreal delivery of rAAV-2/2.CMV.gfp in dog (DIV1). The PCR analysis was carried out as described. The DNA marker (M), positive control on 25 pg of vector plasmid (+), and negative control on water (−) are included. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
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Fig. 5 Biodistribution of vector sequences in the brain following intravitreal injection of rAAV-2/2.CMV.gfp in dogs. The visual pathway is represented. The stars indicate detection of vector sequences in DIV1, DIV2, or DIV3. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
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