Figure 2

Episomal ssODN-mediated gene repair assay. (A) Dose-curve for molar ratios of plasmid/Luc PO AS 45-mer. Total protein content (black diamonds) is shown on the right y-axis. (B) HEK293T cells were transfected with both, Luc targeting oligonucleotides and the repair target vector peGFPLucMut using the molar ratio plasmid/gene repair agent of 1/100. Two days after, cell samples were submitted to luciferase activity measurement to quantify the gene repair efficiency (left y-axis). Total protein content is indicated with the black squares. (C) Similar experiments as in (B) were conducted with the eGFP targeted oligonucleotides and the mutated pmeGFP using the molar ratio plasmid/gene repair agent of 1/100. The repair was followed by eGFP positive cells counting using flow cytometry. Luc, luciferase; GFP, green fluorescent protein; AS, antisense; S, sense; PO, phosphodiester; PS, phosphorothioate; LNA, locked nucleic acid; 35, 35mer; 45, 45mer; Ctr, non-correcting control oligonucleotide; Rep, repair oligonucleotide.