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Split intein vectors permit oversize FLIM-FRET biosensor neuronal expression

Lee, J. O.; Puhl, H.; Holder, A.; Sheridan, A.; Darden, C.; Shah, M.; Nguyen, T.; Hines, K.; Augustin, S. M.; Kim, Y.; Vogel, S. S.; Lovinger, D. M.

2026-06-05 bioengineering
10.64898/2026.06.02.725428 bioRxiv
Show abstract

Adeno-associated virus (AAV) packaging limits constrain the design, performance, and in vivo application of genetically encoded biosensors. We developed a split intein-mediated reconstitution strategy enabling modular delivery and reassembly of oversized fluorescence lifetime-based biosensors. Using this platform, we engineered an oversized cAMP sensor compatible with one-photon fluorescence lifetime measurements, enabling monitoring of intracellular signaling dynamics in distinct neuronal subtypes in freely behaving mice.

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