AAV9-Mediated Expression of AP2-mCherry

An AAV serotype 9 was produced for expression of the µ2-subunit of the AP2 clathrin adaptor tagged with mCherry (AP2-mCherry) (Taylor et al., 2011 (link)). AAV2/9 pseudotyped vectors were prepared by tri-transfection in 293 cells as described previously (Rivière et al., 2006 (link)) using the pSMD2-AP2-mCherry plasmid, pXX6 plasmid coding for the viral sequences essential for AAV production, and the pRepCap plasmid coding for serotype 9 capsid. Vector particles were purified on iodixanol gradient and concentrated on Amicon Ultra-15 100K columns (Merck-Millipore, USA). The viral genomes titer (vg/ml) was determined by quantitative real-time PCR. WT and HTZ KI-Dnm2^R465W mice were injected at 6 mo of age. One intramuscular injection (40 µl/TA) of AAV9-AP2-mCherry at 6.10¹¹ vg/ml was performed in TA muscles using a 29G needle.