Figure 1

Reporter constructs and correction agents. (A) Representation of the peGFPLucMut plasmid. A premature stop codon (black plain bar) was introduced into the luciferase portion of the eGFP-Luciferase fusion gene driven by a CMV promoter. (B) In the pmeGFP plasmid, the eGFP gene harbors a premature stop codon (black plain bar) and a silent mutation (white bar) used as tag sequence. The following information are given for the repair target vectors: the asterisk indicates the position of the mutation to correct and the half-arrows represent the location of the primers used to produce the dsDNA fragments. The length of homology between the gene repair agents and the targeted sequences are represented below each vector. Sequence of the primers used to generate the dsDNA fragments were as follows: F2200, 5'-AAATGTCGTAACAACTCCGCC-3'; F1000, 5'-TACAACTACAACAGCCACAAC-3'; F500, 5'-CGCCAAAAACATAAAGAAAGG-3'; F200, 5'-GCTATGAAGAGATACGCCCT-3'; R2200, 5'-AATGTAGCCATCCATCCTTGTC-3'; R1000, 5'-AATCTCACGCAGGCAGTTC-3'; R500, 5'-CGAACGTGTACATCGACTG-3'; R200, 5'-CAACACCGGCATAAAGAATTG-3' and R700, 5'-TGCTCAGGTAGTGGTTGTCG-3. CMV, immediate-early cytomegalovirus promoter (white box); eGFP gene (light grey box); Luciferase gene (dark grey box); pA, SV40 early mRNA polyadenylation box. BglII and XbaI, location of the BglII and XbaI restriction sites. (C) Luciferase activity and transfection efficiency expressed as a percentage obtained with HEK293T cells using different plasmids.