Comparative Analysis of DNA Nanoparticles and AAVs for Ocular Gene Delivery
Figure 5
Subretinal injection of AAVs but not NPs leads to GFP expression in the brain.
P30 Balb/C mice were subretinally injected bilaterally with NP-CBA-GFP, NP-MOP-GFP (6.911 vg), AAV2-CBA-GFP, or AAV5-MOP-GFP (109 vg). a. Native GFP fluorescence in the optic nerve (ON) was assessed by examination of central retinal cryosections. b. Whole brain schematic showing approximate regions where slices shown in (c), are captured. c. Transverse cryosections of whole brain were prepared for confocal microscopy at PI-90 days. Shown are representative low magnification (top row) and higher magnification (bottom row-magnification of box) images of native GFP fluorescence in the visual tract in animals injected with AAV2-CBA-GFP. d. GFP vector DNA was amplified from genomic DNA harvested from the eye, ON, optic chiasm, lateral geniculate nucleus (LGN), and visual cortex (VC). Shown are representative agarose gels from samples analyzed at PI-30 and PI-60 days. N = 8 (eye, ON, LGN, VC) and 4 (chiasm). cp: cerebral crus; InG: layers of superior colliculus; Op: optic nerve layer of the superior colliculus; opt: optic tract; ox: optic chasim; sox: supraoptic decussation; so: supraoptic. Scale bars b. 40 µm, d. 600 µm (top) and 160 µm (bottom). N values for a, b and d can be found in Table 1.