Optogenetics
rAAV-EF1α-Con Fon hChR2(H134R)-EYFP-WPRE-hGH polyA
The broad-spectrum promoter EF1α regulates the expression of light-activated elements and the expression of ChR2-EYFP when Cre and Flp are present at the same time. This AAV expresses hChR2(H134R)-EYFP driven by an ubiquitous Ef1α promoter. The 1.2 Kb EF1α promoter is a commonly used, constitutive promoter derived from human elongation factor-1 alpha (EF1α). It is often used in conditions where the other constitutive promoters, such as CMV, have diminished activity or have been silenced. 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. The wild-type, as well as a few mutations, provide the fastest excitation of the channelrhodopsins offered, and are widely used in optogenetics techniques in neuroscience. Due to delayed channel closure, hChR2(H134R) is a gain-of-function mutation that produces larger photocurrents than wild-type hChR2, but slows down channel kinetics.
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
hChR2(H134R)
EF1α Broad-spectrum Promoter
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
hChR2(H134R): It is the most common form of ChR2. h refers to the coding sequence of the gene that has been optimized for the human codon table. H134R refers to the mutation of histidine at position 134 to arginine. This mutation inhibits the binding of H+ by ChR2 and increases the passing ability of Na+, thereby increasing the overall conductivity of the molecule and thereby increasing the inward current excited by light. Generally, blue light around 470nm is used for excitation.
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
Con Fon: When Cre and Flp recombinase are present at the same time, the target gene can be expressed normally.
Con Fon: Only when Cre and Flp recombinase are present at the same time can the target gene 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
Boolean Algebra
