Figure 8
From: Modeling and Preventing Progressive Hearing Loss in Usher Syndrome III
AAV-mediated Clrn1-HA gene expression in hair cells of the organ of Corti in wild-type mice. High titer stock (>1013 vg/ml) of AAV8-Clrn1-HA was injected through the RWM of the wild-type mice at P1-P3, and the organ of Corti was examined for HA expression in the cochlea whole mount at P10 by immunofluorescence using anti-HA antibody. (A) Schematic representation of the AAV vector construct used for immunolocalization. The AAV expression cassette contains the mouse Clrn1 coding sequence (CDS), without its UTR sequences, fused to the CDS for HA epitope. Serotype 8 AAV capsid was used to package this construct. The ubiquitous small CBA promoter drives expression of Clrn1-HA in the vector. The expression cassette is flanked by inverted terminal repeats (TR) of AAV serotype 2; poly-A, SV40 polyadenylation signal. Immunolocalization of CLRN1-HA in the organ of Corti of transfected mice using antibodies to the HA epitope. Representative specimens from the mid-basal turn of the cochlea from 2 of the 5 mice injected with AAV8-Clrn1-HA are shown in the left top (B) and bottom (C) panels. Cross sections of the cochlea were examined to determine whether the AAV8-mediated transfection was restricted to the hair cells (panels D and E).
