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Chemogenetics

rAAV-CAG-DIO-hM3D(Gq)-mCherry-WPRE-hGH polyA

The broad-spectrum promoter CAG regulates the expression of Cre-dependent chemical genetic activation elements. hM3D(Gq)-mCherry is expressed in the presence of Cre, but hM3D(Gq)-mCherry is not expressed in the absence of Cre. hM3D(Gq) is another modified human-derived G protein-coupled receptor. After activation with clozapine N-oxide (CNO), it activates the Gq signaling pathway to inhibit cells.

Product No.CS-1738
Serotype

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

Receptor

hM3D(Gq)

Promoter

CAG Broad-spectrum Promoter

Storage

Store at -80°C

Volume
1E+12GC
1E+12GC>1E+14GC
Titer
≥1E+13GC/mL
≥1E+13GC/mL≥5E+12GC/mL
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Delivery Information

Usually Ships in 7 Business Days;

Product Details

Promoter Details

CAG, strong promoter, an artificially constructed combined promoter, composed of cytomegalovirus early enhancer and chicken β-actin promoter, 1731bp.

Functional Gene Details

hM3D(Gq): It is an artificial receptor modified from the human muscarinic acetylcholine receptor M3 that only responds to CNO and is no longer activated by acetylcholine. In some neurons, after CNO binds hM3D(Gq), it activates the Gq protein-coupled phosphatase PLCβ, causing PIP2 (phosphatidylinositol(4,5)bisphosphate, phosphatidylinositol diphosphate) to be degraded, thereby opening the KCNQ outward potassium ion channel closed by PIP2, so that the cell membrane is depolarized, forming an action potential, that is, activating neurons. In neurons that do not express KCNQ or Gq, the function of hM3D(Gq) may be different.

Fluorescent Tag

mCherry

Fluorescent Protein Details

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

Virus Type

adeno-associated virus

Application

Chemical genetics technology/DREADDS (designer receptors exclusively activated by designer drugs) technology can be used to regulate the activities of neurons or other types of cells. For example, by injecting the virus into a specific brain area of ​​a mouse, and after hM3D(Gq) is expressed in the mouse, CNO (clozapine-N-oxide, N-oxide clozapine) is given via intraperitoneal injection or diet, which can activate neurons over a long period of time (on the order of hours). DREADDs technology can be used to explore neurons and neural circuits related to memory formation, food intake regulation and arousal.

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