PFD-rAAV-EF1a-axon-hChR2(E123T/T159C)-mCherry-WPRE-hGH polyA

Promoter Details

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

Functional Gene Details

hChR2(E123T/T159C): It is called one of the upgraded versions of ChETA in Karl Deisseroth's laboratory. By introducing the point mutation of T159C, the shortcoming of ChETA's weak inward current has been significantly improved. The inward current in the plateau period is close to hChR2(H134R), while also maintaining the rapid closing characteristics of ChETA molecules, giving the channel excellent properties. Based on the E123T mutation, T159C has a significantly better effect on increasing inward current than H134R. The excitation light wavelength is 473nm, suitable for high-frequency light activation.

Fluorescent Tag

mCherry

Fluorescent Protein Details

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

Virus Type

adeno-associated virus

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.

Localization Element Details

axon: Axonal localization.

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.