Figure 3 | Gene Therapy

Figure 3

From: Development of an optimized AAV2/5 gene therapy vector for Leber congenital amaurosis owing to defects in RPE65

Figure 3

In vitro RPE65 protein production following transfection of human (293 T) and murine (3T3) cells with plasmids carrying wild-type hRPE65 cDNA or codon-optimized hRPE65 cDNA with SV40 intron. Western blot of whole cell lysates following transfection of pD10/CMV.hRPE65 plasmid (hRPE65) or pD10/CMV.OPTIRPE65 (OPTIRPE65) plasmid in human and mouse cells. 293 T and 3T3 cell lines were transfected with equal amounts of either pD10/CMV.hRPE65 or pD10/CMV.OPTIRPE65 plasmid and 10 μg of whole cell lysates were used for immunoblotting with a human RPE65 antibody (RPE65). Histone 2B (H2B) was used as a loading control. Quantification of RPE65 protein band intensities and comparison between hRPE65 and OPTIRPE65 for both 293 T and 3T3 cell lines. (P=0.04; mean±s.d.; n=2).

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