1. Improved Retinal Transduction In Vivo and Photoreceptor-specific Transgene Expression Using Adenovirus Vectors With Modified Penton Base 
Siobhan M Cashman, Laura McCullough, Rajendra Kumar-Singh 
Molecular Therapy 
Volume 15, Issue 9, Pages (September 2007)
DOI: /sj.mt
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Structure and characterization of EGFPNAd5/F17 virus. (a) N terminal amino acid sequences of wild-type Ad5, Ad17, and the Ad5F17 fusion fiber. The hybrid fiber is composed of the first 10 amino acids of Ad5 followed by amino acids 11–366 of wild-type Ad17. (b) Structure of EGFPNAd5/F17. (c) Western blot analysis of protein prepared from purified virions indicating the presence of both the monomeric and the trimerized fiber. (d) GFP expression in the RPE cells (arrow) and occasional Müller cells (arrowhead) in murine retina infected with EGFPNAd5/F17. The ×40 images are the boxed areas in the ×10 images. ITR, adenovirus inverted terminal repeat; Ψ, Ad packaging signal; BGHpA, bovine growth hormone polyadenylation signal; BF, bright field; CMV, cytomegalovirus promoter/enhancer; E, early region; EGFP, enhanced green fluorescent protein; GFP, green fluorescent protein; MLT, major late transcription unit; RPE, retinal pigment epithelium.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
Comparision of cytomegalovirus (CMV) and chicken beta actin (CBA) promoters in Ad5 background in vivo. Low (a–c and g–i, ×10) and high magnification (d–fand j–o, ×40) images from frozen sections of murine retina injected with a virus expressing green fluorescent protein (GFP) from a CMV promoter (Ad5CMVGFP) or a CBA promoter (Ad5CAGGFP). The ×40 images encompass boxed areas (P,Q,R) depicted in the ×10 images. GFP-positive photoreceptors (PRs) are present (k) in areas with GFP-positive RPE but absent at the edge of the injection site (n). Sporadic GFP-positive ganglion cells in (k) demarcate the inner edge of the retina and ganglion cell layer, more clearly seen in the merged image (l). BF, bright field; ONL, outer nuclear layer; RPE, retinal pigment epithelium. Exposure k and n = 2 seconds).
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Binding of RGD-TRITC or RGE-TRITC to C57BL/6J mouse retina in vivo 90 minutes after subretinal injection. (a) RGD-TRITC peptide primarily binds the RPE (arrow) and blood vessels (arrowheads) and does not substantially bind the photoreceptors (PRs). (b) Control RGE-TRITC peptide does not bind RPE (arrow) but does bind some blood vessels (arrowheads); also, it does not bind PR cells. BF, bright field; RPE, retinal pigment epithelium; TRITC, tetramethylrhodamine isothiocyanate.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Transduction of photoreceptors (PRs) by Ad5CAGGFPΔRGD. Low power (a–c, ×4) and higher power (d–f, ×20; g–l, ×40) images of mouse retina injected with Ad5CAGGFPΔRGD indicating a large portion of the neural retina to be green fluorescent protein (GFP)-positive (b,c). d–i represent boxed areas M in a–c. j–l represent relatively untransduced area N boxed in a. Exposure for k = 2 seconds, similar to e and h, indicates a lack of autofluorescence as a contributor to GFP-positive cells in e and h. Inset in h indicates a very large number of GFP-positive inner segments (IS). Because of some variability in subretinal injections and sizes of retinal detachment, we observed a variation in the total area of retina transduced by the virus. This image depicts a result found in approximately 50–60% of injected mice. However, PR transduction was observed in almost all injected mice. BF, bright field; GFP, green fluorescent protein; ONH, optic nerve head; ONL, outer nuclear layer; PR, photoreceptor; RPE, retinal pigment epithelium. GCL, ganglion cell layer.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
Photoreceptor (PR)-specific transgene expression from Ad5RhoGFPΔRGD. Frozen retinal sections 3 weeks after injection from mice injected subretinally with Ad5RhoGFPΔRGD demonstrating a spread of GFP (b) that localized exclusively to the outer nuclear layer (e, f), a region that contains the PR cell bodies. ×40 images (d–f) are from a different area of the same retina shown in (a–c). Individual GFP-positive PR outer segments are visible in e. BF, bright field; GCL, ganglion cell layer; GFP, green fluorescent protein; ONL, outer nuclear layer; PR, photoreceptor; RPE, retinal pigment epithelium.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions