AAV with CAG promoter driven eArchT3.0-mCherry

This AAV expresses eArchT3.0-mCherry driven by an ubiquitous CAG promoter.

The roughly 1.8Kb CAG promoter (also known as CBA promoter or CAGGS promoter) is a strong synthetic promoter frequently used to drive high levels of gene expression in mammalian cells. The CAG promoter is composed of the following regulatory elements: (C) cytomegalovirus (CMV) early enhancer element; (A) the promoter region, the first exon, and the first intron of chicken beta-Actin gene, and (G) the splice acceptor of the rabbit beta-Globin gene. Like the EF1a promoter, the CAG promoter is commonly used as an alternative for the CMV promoter, from which the expression is decreased due to methylation/silencing. In many cell types tested, the CAG and EF1a promoters give much higher levels of expression than other commonly used cellular promoters such as the UBC and PGK promoters.

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.