Figure 3: Full-length dystrophin delivery with template-switching vector.
From: Lentiviral vectors can be used for full-length dystrophin gene therapy
(a) Schematic for the modified dystrophin vector. DNA synthesis must initiate on the 5′ strand and strand transfer to the 3′ LTR will then permit synthesis of a functional provirus following template-switching within the region of dystrophin homology (shaded). A functional provirus can only be produced following recombination because essential elements are split between both strands. (b) Analysis of dystrophin-GFP expression in HEK 293T cells at 4 days post-transduction. Mock-transduced HEK 293T cells were used to set the baseline of GFP fluorescence. Bars represent average GFP readings and standard deviation. All samples are N = 6. **P < 0.01 by Mann-Whitney U test. (c) Nested PCR for full-length dystrophin-GFP from transduced HEK 293T genomic DNA. Products separated on 1% agarose gel show the presence of a 6.2 kb band in sample transduced with the heterozygous vector. L, 10 kb+ ladder; 1, water; 2, mock-transduced HEK 293T; 3, homozygous-transduced HEK 293T; 4, heterozygous-transduced HEK 293T.
