PurposeDouble floxed genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production vector under the control of the mammalian promoter (EF1a)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||179460||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV1||179460-AAV1||Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid.||$380|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5612
- Total vector size (bp) 6899
Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Insert Size (bp)1287
- Promoter EF1a
- Cloning method Restriction Enzyme
- 5′ cloning site Bsp1407I (not destroyed)
- 3′ cloning site NheI (not destroyed)
- 5′ sequencing primer TCAAGCCTCAGACAGTGGTTC
- 3′ sequencing primer GGTTGATTATCGATAAGCTTGATATCG (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Information for AAV1 (Catalog # 179460-AAV1) ( Back to top )
Ready-to-use AAV1 particles produced from pAAV-EF1a-DIO-JEDI-2P-WPRE (#179460). In addition to the viral particles, you will also receive purified pAAV-EF1a-DIO-JEDI-2P-WPRE plasmid DNA.EF1a-driven, Cre-dependent expression of voltage indicator JEDI-2P. These AAV preparations are suitable purity for injection into animals.
- Volume 100 µL
- Titer ≥ 1×10¹³ vg/mL
- Pricing $350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- Storage Store at -80℃. Thaw just before use and keep on ice.
- Shipment Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmids encode adenoviral helper sequences and AAV rep gene, AAV1 cap gene
- Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
- Serotype AAV1
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene cEGFP (Cre-dependent)
Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). The specific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for more information.
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.1-0.8% of viral particles in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, it is necessary to optimize the injection volume and viral titer to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral particle dosage in order to reduce the likelihood of Cre-independent expression.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pAAV-EF1a-DIO-JEDI-2P-WPRE was a gift from Francois St-Pierre (Addgene plasmid # 179460 ; http://n2t.net/addgene:179460 ; RRID:Addgene_179460)
For viral preps, please replace (Addgene plasmid # 179460) in the above sentence with: (Addgene viral prep # 179460-AAV1)
For your References section:Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy. Liu Z, Lu X, Villette V, Gou Y, Colbert KL, Lai S, Guan S, Land MA, Lee J, Assefa T, Zollinger DR, Korympidou MM, Vlasits AL, Pang MM, Su S, Cai C, Froudarakis E, Zhou N, Patel SS, Smith CL, Ayon A, Bizouard P, Bradley J, Franke K, Clandinin TR, Giovannucci A, Tolias AS, Reimer J, Dieudonne S, St-Pierre F. Cell. 2022 Aug 16. pii: S0092-8674(22)00916-3. doi: 10.1016/j.cell.2022.07.013. 10.1016/j.cell.2022.07.013 PubMed 35985322