Fig. 1: ATAC-seq identifies candidate GREs to restrict viral GJB2 expression.

a Overview of the experimental design. ATAC-seq Assay for Transposase-Accessible Chromatin with Sequencing, AAV Adeno-associated virus, ABR Auditory Brainstem Response, DPOAE Distortion Product Otoacoustic Emissions. b Identification of potential GREs with ATAC-seq. Ten cochleas from neonatal mice age P2 (n = 3), P5 (n = 3), P8 (n = 4) were harvested and processed for ATAC-seq. Peaks representing open chromatin are shown for a ~300 kb region of mouse genome near the Gjb2 locus. Peaks for a neuronal sample are shown at the top; sequence conservation among 42 mammalian genomes is plotted at the bottom. c ATAC-seq peaks at the Gjb2 gene and elements chosen for cell-specific vectors. The human or mouse coding sequence (green) was preceded by a conserved region that includes the human equivalent of mouse exon 1 (violet; GRE1) and followed by the human non-coding segment of exon 2 (violet; GRE2). d Schematics of the AAV vectors used in the study. All vectors are single stranded. ITR inverted terminal repeat, CBA hybrid CMV enhancer/chicken β-actin promoter, HA hemagglutinin tag, GRE1 gene regulatory element 1, GRE2 gene regulatory element 2, WPRE woodchuck hepatitis virus post-transcriptional regulatory element, CDS Coding DNA Sequence, BGH bovine growth hormone polyadenylation signal.