FIGURE 4.

Sequence analysis of the edited MECP2 gene in hemizygous male S134C Rett syndrome cells. (A) Editing of the MECP2 gene in S134C cells. Shown are the mutant S134C genome and the AAVHSC-226 editing vector. Also shown are: i) positions of SNPs which differed between the editing vector and the S134C genome, ii) location of linkers L1 and L2, iii) forward and reverse primers, iv) the S134C mutation. The primers used for TI analyses are depicted as thick red arrows. Primers 1F and 3R are specific for chromosomal sequences external to the homology arm. Primer 2R is specific for linker L2. Positions for HR crossover events between the mutant genome and the editing vector genome were identified by the presence or absence of markers in the edited genome and are depicted. (B) Observed 5′ editing outcomes. Two editing outcomes identified at the 5′ end based on the SNPs and linker sequences as markers are shown. (C) Observed 3′ editing outcomes. Two editing outcomes identified at the 3′ end based on the SNPs and linker sequences as markers are shown.