Self-complementary AAV5 Vector Facilitates Quicker Transgene Expression in Photoreceptor and Retinal Pigment Epithelial Cells of Normal Mouse

摘要: To clarify whether transduction efficiency and cell type specificity of self-complementary (sc) AAV5 vectors are similar to those standard, single-stranded in normal retina, one micro liter scAAV5-smCBA-GFP vector (1 x 10(12) genome-containing particles/ml) AAV5-smCBA-GFP were subretinally or intravitreally (in both cases through the cornea) injected into right left eyes adult C57BL/6J mice, respectively. On post-injection day (PID) 1, 2, 5, 7, 10, 14, 21, 28 35, enucleated; retinal pigment epithelium (RPE) wholemounts, neuroretinal wholemounts eyecup sections prepared evaluate green fluorescent protein (GFP) expression by microscopy. GFP following trans-cornea subretinal injection was first detected RPE around PID 1 between 2 5; peaked stabilized 10-14 P14 P21 with strong, homogeneous fluorescence covering entire wholemounts. The frozen supported findings from wholemounts: appeared 1-2 soon spread photoreceptors (PR) cells; moderate found mainly PR layers; 14 strong homogenous observed cells. 5-7 7-10; ssAAV5-mediated at 21 wholemounts; treated also obtained then Peak mediated scAAV5 that AAV5. However, peak cells stronger than No any (RPE sections) after intravitreal delivery either scAAV5-GFP AAV5-GFP. Neither nor can transduce injection. used this study directs an earlier onset transgene matched vector, has Our data confirm previous reports scAAV have single strand AAV (Natkunarajah et al., 2008; Yokoi 2007). may be more useful when addressing certain and/or cell-related models dystrophy, particularly for mouse human retinitis pigmentosa require rapid robust prevent early degeneration