PurposeCre-dependent AAV expression of humanized ChR2 (with H134R mutation) fused to eYFP for optogenetic activation
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||127090||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV PHP.eB||127090-PHPeB||Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid.||$380|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression, AAV
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Insert Size (bp)1662
MutationHumanized ChR2 gene with histidine 134 changed to arginine (H134R), to achieve higher currents
- Promoter CAG
/ Fusion Protein
- eYFP (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site MluI (not destroyed)
- 3′ cloning site KpnI (not destroyed)
- 5′ sequencing primer n/a
- 3′ sequencing primer n/a (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byKarl Deisseroth* (see below)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
*Note: This construct is based on Addgene ID 20298 (the Ef1a promoter was changed to the CAG promoter).
Information for AAV PHP.eB (Catalog # 127090-PHPeB) ( Back to top )
Ready-to-use AAV PHP.eB particles produced from pAAV-CAG-DIO-ChR2(H134R)-eYFP (#127090). In addition to the viral particles, you will also receive purified pAAV-CAG-DIO-ChR2(H134R)-eYFP plasmid DNA.CAG-driven, Cre-dependent expression of humanized channelrhodopsin H134R mutant fused to EYFP, for optogenetic activation. These AAV were produced with the PHPeB serotype, which permits efficient transduction of the central nervous system. 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
encode adenoviral helper sequences and AAV rep gene, PHP.eB cap gene
pUCmini-iCAP-PHP.eB (plasmid #103005)
- Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
- Serotype PHPeB (plasmid #103005)
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene EYFP (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
Terms and Licenses
- Not Available to Industry
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.
Please note that this vector may have a higher level of recombination than our other FLEX vectors. We are making it available because it is still useful to some scientists, but understand that you may see expression in the absence of Cre.
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.Citation Information: When using the PHP.eB serotype in future publications, please acknowledge Viviana Gradinaru and cite Chan et al., Nat Neurosci, 20(8):1172-1179. Pubmed.
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-DIO-ChR2(H134R)-eYFP was a gift from Viviana Gradinaru (Addgene plasmid # 127090 ; http://n2t.net/addgene:127090 ; RRID:Addgene_127090)
For viral preps, please replace (Addgene plasmid # 127090) in the above sentence with: (Addgene viral prep # 127090-PHPeB)
For your References section:Identification of peripheral neural circuits that regulate heart rate using optogenetic and viral vector strategies. Rajendran PS, Challis RC, Fowlkes CC, Hanna P, Tompkins JD, Jordan MC, Hiyari S, Gabris-Weber BA, Greenbaum A, Chan KY, Deverman BE, Munzberg H, Ardell JL, Salama G, Gradinaru V, Shivkumar K. Nat Commun. 2019 Apr 26;10(1):1944. doi: 10.1038/s41467-019-09770-1. 10.1038/s41467-019-09770-1 PubMed 31028266