Cell type specific targeting of nicotinic receptor activity with tethered ligand viral vectors.

Ch produced strong, long-lasting, and E61C-dependent suppression of ACh-evoked currents, with near-complete blockade following prolonged ligand exposure. The system performed robustly in both DAT-Cre mice and rats receiving co-infused AAV-Cre, establishing effective cross-species use. From our results, we infer that any heteromeric receptor containing ≥1 engineered β2 subunit becomes ligand-sensitive, and that viral expression efficiently converts the endogenous β2 receptor pool without increasing total receptor abundance. This AAV-based, Cre-dependent E61C system provides a practical, optics-free, and species-flexible platform for cell-type-specific silencing of β2-containing nAChRs. Its compatibility with widely used Cre lines and suitability for rat behavioral paradigms-including nicotine self-administration and sustained-attention tasks-positions it as a versatile tool for dissecting nicotinic signaling in intact neural circuits.