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Quantitative comparison of unmodified and capsid mutated AAV vectors in vivo.

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posted on 26.04.2013, 00:39 by Christine N. Kay, Renee C. Ryals, George V. Aslanidi, Seok Hong Min, Qing Ruan, Jingfen Sun, Frank M. Dyka, Daniel Kasuga, Andrea E. Ayala, Kim Van Vliet, Mavis Agbandje-McKenna, William W. Hauswirth, Sanford L. Boye, Shannon E. Boye

Transduction efficiency of unmodified and capsid-mutated scAAV2, scAAV5 and scAAV8 vectors in Rho-GFP mice. FACS analysis was used to quantify the percentage of cells that were GFP positive (PRs), mCherry positive (any retinal cells transduced with AAV) and both GFP and mCherry positive (PRs transduced by AAV). Representative plots for a negative control (uninjected retina) and 2 pooled retinas injected with scAAV2(quadY-F+T-V) are shown in Panels A and B, respectively. Cells that were both GFP and mCherry positive are shown in the top right of Panels A and B and represent the percent of transduced PRs. The bottom right of Panels A and B show cells that were mCherry positive but GFP negative, representing off-target transduction. The percentage of mCherry positive PRs (a measure of in vivo PR transduction efficiency for each vector) in retinas injected with unmodified or capsid-mutated scAAV vectors is shown in Panel C.

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