Optogenetics
rAAV-EF1α-DIO-ChETA-EYFP-WPRE-hGH polyA
The broad-spectrum promoter EF1α regulates the expression of Cre-dependent light-activated elements. ChETA-EYFP is expressed when Cre is present, but ChETA-EYFP is not expressed when Cre is absent. This AAV expresses ChETA-EGFP driven by a neuron hSyn promoter. hSyn is a Human synapsin 1 gene promoter. Channelrhodopsins are light-gated ion channels that exist naturally in microalgae. hChR2 is a humanized version of ChR2 for mammalian expression. It is maximally excited by 470 nm light. ChETA is a mutant of ChR2, combined with E123T and H134R mutations, has a faster dynamic change. ChETA neurons allows to emit a 200Hz spike when stimulated by a laser, while others ChR2 can only reach 40Hz.
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, Please consult for more serotypes
ChETA
EF1α Broad-spectrum Promoter
Store at -80°C
Delivery Information
Usually Ships in 7 Business Days；
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
ChETA: There is a point mutation of E123T, which accelerates the closing speed of the ion channel after light withdrawal, making the channel more sensitive to changes in light signals. There is also a mutation of hChR2 (H134R) to increase the inward current. Experiments have shown that ChETA can excite action potentials at a frequency of 200Hz in some neurons, and it is a commonly used high-frequency excitation tool. Compared with ChR2, its maximum excitation wavelength is red-shifted by about 20nm, but it can generally be excited with blue light of about 470nm. The excitation light wavelength is 490nm, suitable for high-frequency light activation.
Fluorescent Tag
EYFP
Fluorescent Protein Details
EYFP: Yellow fluorescent protein, excitation wavelength 513nm, emission wavelength 527nm.
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.
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
