Chronic neuropathic pain’s impact, persistence, and limited treatments render it relevant for gene therapy. Here, we describe the development and application of self-assembling dimeric peptide inhibitors of the pain-associated scaffolding protein PICK1 (protein interacting with C-kinase 1) delivered by adeno-associated viral (AAV) vectors. In mice, these peptides prevented mechanical allodynia in inflammatory and neuropathic pain models and reversed neuropathic pain for up to one year. Targeting of somatosensory pain pathways relieved pain without overt side effects, while selective transduction of dorsal root ganglion (DRG) neurons was sufficient for providing pain relief. Using proteomic and phosphoproteomic analysis of DRG tissue, we identified protein kinase C alpha (PRKCA) substrate candidate, that potentially shape this pain-relieving phenotype. We also confirmed PICK1 expression and peptide target engagement in mice and human donor tissue, supporting the potential of AAV-based PICK1 inhibitors as a clinically meaningful strategy for neuropathic pain conditions.