PurposeAAV expression of CaMKIIa-driven hChR2 variant for ultrafast optogenetic control.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||35512||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
|AAV9||35512-AAV9||Virus (100 µL at titer ≥ 1×10¹³ vg/mL)|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5385
Modifications to backboneAddition of a CaMKIIa promoter and a WPRE
Vector typeMammalian Expression, AAV
Growth in Bacteria
Copy numberHigh Copy
Insert Size (bp)1646
MutationE123T and T159C
- Promoter CaMKIIa
/ Fusion Protein
- mCherry (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer AGTCCTGCAGTATTGTGTAT
- 3′ sequencing primer GCAATAGCATGATACAAAGG (Common Sequencing Primers)
Information for AAV9 (Catalog # 35512-AAV9) ( Back to top )
Ready-to-use AAV9 particles produced from pAAV-CaMKIIa-hChR2(E123T/T159C)-mCherry (#35512). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa-hChR2(E123T/T159C)-mCherry plasmid DNA.CaMKIIa-driven, humanized channelrhodopsin E123T/T159C mutant fused to mCherry for optogenetic activation. 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, AAV9 cap gene
- Buffer PBS + 0.001% Pluronic F-68
- Serotype AAV9
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene mCherry
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.
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-CaMKIIa-hChR2(E123T/T159C)-mCherry was a gift from Karl Deisseroth (Addgene plasmid # 35512 ; http://n2t.net/addgene:35512 ; RRID:Addgene_35512)
For viral preps, please replace (Addgene plasmid # 35512) in the above sentence with: (Addgene viral prep # 35512-AAV9)
For your References section:Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. Mattis J, Tye KM, Ferenczi EA, Ramakrishnan C, O'Shea DJ, Prakash R, Gunaydin LA, Hyun M, Fenno LE, Gradinaru V, Yizhar O, Deisseroth K. Nat Methods. 2011 Dec 18;9(2):159-72. doi: 10.1038/nmeth.1808. 10.1038/nmeth.1808 PubMed 22179551