PFD-rAAV-EF1a-DIO-eArch3.0-EYFP-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

eArch3.0, enhanced light-driven outward hydrogen ion pump, is an engineered variant of Arch (Archaerhodopsin-3). After light activation, hydrogen ions can be pumped from intracellular to extracellular, thereby inducing hyperpolarization of neurons and inhibiting electrical activity. The excitation wavelength is approximately 566 nm. Compared with the original Arch, eArch3.0 achieves higher expression efficiency by optimizing membrane positioning, resulting in stronger photocurrent at the same expression level.

Fluorescent Tag

EYFP

Fluorescent Protein Details

EYFP: Yellow fluorescent protein, excitation wavelength 513nm, emission wavelength 527nm.

Application

Expressing light-sensitive ion channel proteins, such as eNpHR3.0, Arch, etc., in neurons through rAAV viral vectors can specifically inhibit 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.