Chemogenetics
rAAV-EF1a-DIO-hM3D(Gq)-EGFP-WPREs
The broad-spectrum promoter EF1a regulates the expression of Cre-dependent chemical genetic activation elements. hM3D(Gq)-EGFP is expressed in the presence of Cre, but hM3D(Gq)-EGFP 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.
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
hM3D(Gq)
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 in some cells where the CMV promoter is silenced, such as stem cells, 1178bp. Functional gene: 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 the degradation of PIP2 (phosphatidylinositol(4,5)bisphosphate, phosphatidylinositol diphosphate), thereby opening the KCNQ outward potassium channel closed by PIP2, so that the cell membrane is depolarized, forming an action potential, that is, activating the neuron. The function of hM3D(Gq) may be different in neurons that do not express KCNQ or Gq.
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
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
EGFP
Fluorescent Protein Details
EGFP: Green fluorescent protein, excitation wavelength 488nm, emission wavelength 507nm.
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 hM 3 D(G q ) 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. Only in the presence of Cre can the target gene be expressed normally.
Notes
1. Short-term storage at 4°C; Long-term storage at -80°C
2. Can be stored at -80°C for more than 6 months. Each freeze-thaw will reduce the titer 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 experimental needs, the virus can be appropriately diluted and divided into the volume required for each experiment.
Expression and Regulation Mode
Cre-dependent
