Herpes Simplex Virus 1 and Adeno Associated Virus distribution in neurons of the murine trigeminal and superior cervical ganglia

After initial infection at mucosa, herpes simplex virus (HSV) establishes lifelong latency in neurons of the peripheral nervous system, which represents the source of recurrent disease. Current antiviral therapies reduce symptoms and viral shedding, but do not cure the infection. In contrast, gene editing offers the possibility to lethally mutate or even eliminate latent viral genomes. Delivery of gene editing enzymes by Adeno Associated Virus (AAV) vectors represents a promising approach to functionally curing HSV infection. In order to optimize in vivo gene therapy approaches it is necessary to understand which neuronal subtypes within peripheral ganglia are infected by HSV and which subtypes are efficiently targeted by various AAV serotypes. Here we use single cell RNA sequencing (scRNA-seq) to identify neurons expressing HSV genes as well as reporter genes for AAV1, AAV8, AAV-PhP.s, and AAV-Rh10 serotypes. Overall design: 12 adult female Swiss-Webster mice were infected with 10^5 PFU HSV syn17+ by corneal scarification. 3 mice were transduced with AAV1-mScarlet via intradermal injection into the wisker pad; 3 mice were transduced with AAV1-mScarlet via retro orbital IV injection. 3 mice were transduced with AAV8-mEGFP intravenously via retro-orbital injection; 3 mice were transduced with AAV-PHP.S-DsRed-Express2 intravenously via retro-orbital injection; and 3 mice were transduced with AAV-Rh10-mTagBFP-2 intravenously via retro-orbital injection. Trigeminal Ganglia were dissected out of each animal and pooled. Superior Cervical Ganglia were dissected out of each animal and pooled.