AAV with CamKII promoter driven Cre Inducible hChR2(E123A)-Venus

This AAV expresses DIO-hChR2(E123A)-Venus driven by a neuron CamKII promoter.

This 1.3 Kb promoter is derived from the murine α-Calcium/calmodulin-dependent kinase II (CaMKII), a gene with expression restricted to excitatory neurons in the neocortex and hippocampus. In cortical neurons, it was shown that the promoter activity of CaMKII(1.3) is about double that of CaMKII(2.4), probably due to some inhibitory element(s).

Channelrhodopsins are light-gated ion channels that exist naturally in microalgae. 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. hChR2(E123A) mutant accelerates the open state decay but reduces photocurrent amplitudes. Its maximal spectrum is near 485nM.

In the DIO scenario, the transgene of interest is inserted in reverse orientation relative to the 5′ promoter and is flanked by oppositely oriented loxP and lox2272 sites. In the absence of Cre expression, the transgene will not be produced. In the presence of Cre expression, the transgene will be “FLip-EXchanged” or FLEXed, leading to expression of the transgene. This is due to a permanent Cre-mediated recombination/inversion of the flanked transgene. This arrangement is called DIO (double-floxed inverse ORF), Cre-ON, Flex-rev (reverse), Flex-ON/FlexON, or DIO-AAV/AAV-DIO (double-floxed inverse ORF in AAV).