Optogenetics
PFD-rAAV-ChAT-DIO-hChR2(H134R)-EGFP-WPREs
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, AAV3, AAV4, AAV5, AAV6, AAV8, AAV9, retro, AAV11, AAV13, PHP.eB, BR1, B10, MG1.AAV2, olig001, DJ, php.s, 9/retro
hChR2(H134R)
ChAT Neuron-specific Promoter
Store at -80°C
Delivery Information
Usually Ships in 7 Business Days；
Product Details
Promoter Details
ChAT, mouse cholinergic neuron-specific promoter, 1474bp.
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 passage 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
EGFP
Fluorescent Protein Details
EGFP: Green fluorescent protein, excitation wavelength 488nm, emission wavelength 507nm.
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
