AAV with CAG promoter driven hChR2(C128T)-EYFP

This AAV expresses hChR2(C128T)-EYFP 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.

hChR2 is a humanized version of ChR2 for mammalian expression. Wild-type ChR2, as well as a few of its mutations, provides the fastest excitation of the channelrhodopsins offered, and is widely used in in optogenetics techniques in neuroscience. Mutations at C128 (C128A, C128S, C128T) can extend depolarization up to seconds or even minutes; they are sometimes referred to as step-function opsins or SFOs. hChR2(C128T) mutant provides for a 2-second open-state lifetime. It has showed strong expression in the hippocampal neurons and Pichia pastoris. It becomes inactivated at strong light due to a reaction into a deprotonated species.