AAV with CAG2 promoter driven eArchT3.0-mCitrine

This AAV expresses eArchT3.0-mCitrine driven by an ubiquitous CAG2 promoter.

The CAG2 promoter is derived from the commonly used 1.8kb CAG/CBA (also called CAGGS) promoter. To increase the cloning capacity of size-limiting vectors using the CAG promoter, such as AAV or lentiviral vectors, CAG2 was developed by deleting a portion of about 0.6Kb of the chicken beta-actin intron from the original CAG promoter.

Arch and ArchT are light-driven proton pumps found in archaebacteria. They provide silencing via hyperpolarization. Only slightly slower than NpHR2, their peak activation falls at 566 nm. ArchT has about a three-fold higher light sensitivity compared to Arch leading to a similar level of inhibition but increased volume of tissue that will be inhibited.

The trafficking signal (KSRITSEGEYIPLDQIDINV) is also from the inward rectifier potassium channel Kir2.1; it serves to dramatically reduce intracellular accumulation and improve membrane targeting, leading to a profound increase of photocurrents. This is the basis for many third-generation optogenetics tools

This ER export motif (FCYENEV) is from the vertebrate inward rectifier potassium channel Kir2.1. The motif prevents aggregate formation, decreases intracellular accumulations, and enhances tolerability at high expression levels. This is the strategy for developing many second-generation optogenetics tools