FIGURE 7.

Rescue of MECP2 expression in edited hemizygous GM21921 fibroblasts. (A) Schematic showing the r.378_384del mutation in the genome and transcript of GM21921 cells before and after genome editing. The cells contain a deletion at the Intron 3/Exon 4 splice junction resulting in formation of a new splice site and a premature termination codon. AAVHSC7-226 editing restored the wild type splice junction and Exon 4 sequence in r.378_384del cells. The primers used for qRT-PCR are depicted. The forward primer is complementary to the Exon 3/Exon 4 splice junction in the wild-type spliced transcript, which is deleted in GM12921 cells. The reverse primer anneals to Exon 4 downstream of the mutant premature termination codon. (B) Quantitation of restoration of MECP2 expression after editing. Shown is the fold-change in MECP2 transcript levels in AAVHSC7-226 edited GM21921 cells (AAVHSC7-226 Td GM21921) and wild-type AG21802 cells (Untd AG21802) compared with unedited GM21921 cells (Untd GM21921). The fold change represents the average of 2 experiments, with 3 replicates each. Bars represent the standard deviation.