PurposeAAV expression of Cre recombinase-activated jRGECO1b, a red fluorescent calcium sensor protein, from the CAG promoter
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||100855||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV1||100855-AAV1||Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid.|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression, AAV
Growth in Bacteria
Growth Strain(s)NEB Stable
- Promoter CAG
- Cloning method Unknown
- 5′ sequencing primer unknown (Common Sequencing Primers)
This plasmid was previously available as pAAV.CAG.Flex.NES-jRGECO1b.WPRE.SV40 (p3844) from the Penn Vector Core. This plasmid was created as part of the GENIE project at Janelia Research Campus.
Information for AAV1 (Catalog # 100855-AAV1) ( Back to top )
Ready-to-use AAV1 particles produced from pAAV.CAG.Flex.NES-jRGECO1b.WPRE.SV40 (#100855). In addition to the viral particles, you will also receive purified pAAV.CAG.Flex.NES-jRGECO1b.WPRE.SV40 plasmid DNA.CAG-driven, Cre-dependent GECO1b calcium sensor. 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
- Serotype AAV1
- Purification Iodixanol gradient ultracentrifugation
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.CAG.Flex.NES-jRGECO1b.WPRE.SV40 was a gift from Douglas Kim & GENIE Project (Addgene plasmid # 100855 ; http://n2t.net/addgene:100855 ; RRID:Addgene_100855)
For viral preps, please replace (Addgene plasmid # 100855) in the above sentence with: (Addgene viral prep # 100855-AAV1)
For your References section:Sensitive red protein calcium indicators for imaging neural activity. Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman JP, Tsegaye G, Holt GT, Hu A, Walpita D, Patel R, Macklin JJ, Bargmann CI, Ahrens MB, Schreiter ER, Jayaraman V, Looger LL, Svoboda K, Kim DS. Elife. 2016 Mar 24;5. pii: e12727. doi: 10.7554/eLife.12727. 10.7554/eLife.12727 PubMed 27011354