PFD-rAAV-EF1α-DIO-C1V1-mCherry-WPRE-hGH polyA

Promoter Details

Ef1α, strong promoter, human targeted elongation factor 1α (EF1A) gene, can be stably expressed in most types of cells, especially cells where some CMV promoters are silenced, such as stem cells, 1178bp.

Functional Gene Details

C1V1: It is a new ion channel formed by the fusion of ChR1 and VChR1 fragments. This protein is very sensitive. C1V1 expressed in C. elegans can work under 0.2-0.5mW/mm2 light. The maximum absorption wavelength is 540-580nm. It can also be activated by 400nm laser. The common C1V1 carries two point mutations, E122T and E162T, to improve activity, so it is also called C1V1(t/t). The excitation light wavelength is 560nm.

Fluorescent Tag

mCherry

Fluorescent Protein Details

mCherry: red fluorescent protein, excitation wavelength 587nm, emission wavelength 610nm.

Application

Expressing light-sensitive ion channel proteins, such as ChR2, ChrimsonR, etc., in neurons through rAAV viral vectors can specifically excite neurons and explore the effects of light stimulation on neurons, neural circuits and animal behavior.

Regulatory Element Details

DIO: Double-floxed inverse Orientation, is an element that cooperates with Cre recombinase to control the expression of the target gene. If there is no Cre recombinase, the target gene will not be expressed, but in the presence of Cre, the target gene can be expressed normally.

Notes

1. Short-term storage at 4°C; long-term storage at -80°C

2. It can be stored at -80°C for more than 6 months. Each time it is frozen and thawed, the titer will be reduced by 10%, and repeated freezing and thawing should be avoided; if the virus is stored for more than 6 months, it is recommended to re-measure the virus titer before use

3. According to the experimental needs, the virus can be appropriately diluted and divided into the volume required for each experiment.