Figure 1
Construction and characterization of AAV8 (Y733F) vector expressing ACE2 and enhanced green fluorescent protein (eGFP).
(a) Maps of the AAV8(Y733F) vector expressing the human ACE2 gene (hACE2) and a control vector contains coding region for the secreted enhanced green fluorescent protein (eGFP). CBA, CMV-chicken-β-actin promoter; ITR, inverted terminal repeat. (b) Transduction of mouse retina with AAV8 (Y733F) vector expressing eGFP. A single subretinal injection of 1 μl AAV8 (Y733F) vector (1011 vg/ml) resulted in robust expression of eGFP. (i) A retinal whole mount showing the expression of eGFP in the magnification of 100X. (ii) Higher magnification of the same retinal whole mount (200X). (iii) A cross section of a mouse eye showed eGFP expression at three weeks after receiving AAV8(Y733F)-eGFP injection in low magnification of 100X. The eGFP was mainly transducted at the RPE and the PR level of the retina. (iv) Higher magnification of the same cross section (200X). Scale bar = 100 μm. RPE, retinal pigment epithelium; PR, photoreceptor; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; RGC, retinal ganglion cells. (c) The mRNA and protein expression of ACE2 in the eyes injected with AAV8(Y733F)-ACE2 or AAV8(Y733F)-eGFP vectors. Data were shown as mean ± SEM (*p < 0.05, **p < 0.01, n = 4).
