Chemogenetics
rAAV-EF1a-DIO-hM4D(Gi)-EGFP-WPREs
The broad-spectrum promoter EF1α regulates the expression of Cre-dependent chemical genetic repressor elements. hM4D(Gi)-EGFP is expressed in the presence of Cre, but hM4D(Gi)-EGFP is not expressed in the absence of Cre. hM4D(Gi) is another modified human-derived G protein-coupled receptor. After activation with clozapine N-oxide (CNO), it activates the Gi 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
hM4D(Gi)
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
hM4D(Gi): It is an artificial receptor modified from the human muscarinic acetylcholine receptor M4 that only responds to CNO and is no longer activated by acetylcholine. In some neurons, CNO binds to hM4D(Gi) and activates the Gi protein-coupled inward potassium channel GIRK, hyperpolarizing the cell membrane and inhibiting the firing of action potentials in neuronal cells. In cells that do not express GIRK or Gi, hM4D(Gi) functions differently.
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 expressing hM4D(Gi) in the mouse, CNO (clozapine-N-oxide, N-oxide clozapine) can be administered via intraperitoneal injection or diet, which can inhibit 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
