Optogenetics
rAAV-Ef1α-DIO-SwiChRca-TS-EYFP-WPRE-hGH polyA
The broad-spectrum promoter Ef1a regulates the expression of Cre-dependent light repressive elements. SwiChRca-EYFP is expressed in the presence of Cre, but SwiChRca-EYFP is not expressed in the absence of Cre. The C1C2 chimera mutation C128A (SwiChRCA) can slow down the closing of the channel, continuously activate the chloride ion channel by a single blue light, keep the cells in an inhibitory state, and close the chloride ion channel after irradiation with red light.
AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, rh10, DJ, B10, PHP.eB, PHP.S, MaCPNS1/2, retro, anc80l65, AAV3, AAV4, AAV11, AAV13, BR1, MG1.AAV2, olig001, 9/retro, AAV2/2, Please consult for more serotypes
SwiChRca SwiChRCA
EF1α Broad-spectrum Promoter hEF1a
Store at -80°C
Delivery Information
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Product Details
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
SwiChRca: Step-Waveform Inhibitory ChannelRhodopsin is a chloride channel protein modified by Karl's laboratory based on ChR2. It is very sensitive to blue light, and the channel closes very slowly, so short-term light stimulation can continuously activate the channel. Using 632nm red light can speed up the channel closing and end the neuronal inhibition state. The excitation light wavelength is 475nm, and a single light suppression has a long effect.
Fluorescent Tag
EYFP
yellow fluorescent yellow
Fluorescent Protein Details
EYFP: Yellow fluorescent protein, excitation wavelength 513nm, emission wavelength 527nm.
Virus Type
adeno-associated virus
Adeno-associated virus vector Adeno-associated Virus
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.
Expression and Regulation Mode
Cre-dependent
constitutive expression
