AAV with CamKII(0.4) promoter driven C1V1(E122T/E162T)-TS-ECFP

This AAV expresses C1V1(E122T/E162T)-TS-ECFP driven by a neuron CamKII(0.4) promoter.

This 0.4 Kb promoter is derived from murine α-Calcium/calmodulin-dependent kinase II (CaMKII), a gene with expression restricted to excitatory neurons in the neocortex and hippocampus. It was shown that the promoter activity of CaMKII(0.4) is almost identical to that of CaMKII(1.3) in cortical neurons, Compared to CaMKII(1.3), this short promoter allows cloning of longer gene of interest into the AAV construct .

C1V1 is a chimeric channelrhodopsin made to boost membrane expression higher than VChR1 but with a different light spectrum. It is composed of ChR1 and VChR1 fragments, and implements fast, potent optical excitation at red-shifted wavelengths. C1V1(E122T/162T) is highly efficient in neurons, with a maximum absorption at 530nM with a narrow action spectra. Off-kinetics is 35ms.

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