Plasmid and Viral Vector Products

Specificity is determined by the viral capsid proteins, so it is important to select the correct AAV serotype to ensure optimal gene delivery. The table below lists our recommended AAV serotypes for different target tissues. If you are unsure which AAV serotype will work best for your system, we recommend that you try out our AAV GFP Testing Kit. The testing kit contains the following viruses all driven by the strong CMV promoter: GFP AAV1, GFP AAV2, GFP AAV5, GFP AAV6, GFP AAV8, GFP AAV9, and GFP AAV-DJ.

Please use this table to help you decide which promoter may work best for your experiments.

For more information on biosensor gene therapy products, please visit the following Janelia pages about CaMPARI, GCaMP, jRCaMP1/jRGECO1, and iGluSnFR. You can also read our Biocompare article to learn why AAV biosensors are a powerful tool for tracking neuronal dynamics.

The number following GCaMP refers to the generation (3^rd, 5^th, 6^th, or 7^th). The letter at the end refers to the characteristics of the variant.

jGCaMP7s: sensitive and slow
jGCaMP7f: fast kinetics
jGCaMP7b: brighter baseline
jGCaMP7c: high contrast

The plasmids were generated by the Douglas Kim lab. For more information about the jGCaMP7 variants, please see their publication.

Ip, C. W. et al. AAV1/2-induced overexpression of A53T-α-synuclein in the substantia nigra results in degeneration of the nigrostriatal system with Lewy-like pathology and motor impairment: a new mouse model for Parkinson’s disease. Acta Neuropathol Commun 5, 11 (2017).

Koprich, J. B., et al. Towards a Non-Human Primate Model of Alpha-Synucleinopathy for Development of Therapeutics for Parkinson’s Disease: Optimization of AAV1/2 Delivery Parameters to Drive Sustained Expression of Alpha Synuclein and Dopaminergic Degeneration in Macaque. PLoS ONE11, e0167235 (2016).

Koprich, J. B. et al. Progressive neurodegeneration or endogenous compensation in an animal model of Parkinson’s disease produced by decreasing doses of alpha-synuclein. PLoS ONE6, e17698 (2011).

Koprich, J. B., et al. Expression of human A53T alpha-synuclein in the rat substantia nigra using a novel AAV1/2 vector produces a rapidly evolving pathology with protein aggregation, dystrophic neurite architecture and nigrostriatal degeneration with potential to model the pathology of Parkinson’s disease. MolNeurodegener5, 43 (2010)

Yes, all of our AAV Parkinson’s disease tools are replication-incompetent and can be handled safely in a BSL-1 facility.

If you want to generate viral particles, you must not exceed the packaging capacity of the virus. Please note that the packaging capacity refers to the total insert size between and including the ITRs or LTRs.

AAV cloning capacity: 4.7 kb
Adenovirus cloning capacity: 7.5 kb
Lentivirus cloning capacity: 6 kb

The ORF cloned into the pENTER adenovirus shuttle vector can be transferred into the other Charles River’s destination vectors through a simple “cut and paste” restriction enzyme digestion and ligation. A combination of two pairs of restriction enzymes, AsiSI/MluI (96%), AsiSI/RsrII (99%) covers 99.5% of all human cDNA ORFs. All human genes can be covered with a few additional restriction enzyme combinations, such as AsiSI/NotI, AscI/RsrII.

Plasmid miRNA delivery may be sufficient for in vitro work or for conducting pilot studies. However, recombinant viruses are excellent tools for miRNA delivery both in vitro and in vivo. Adenovirus miRNAs in particular are a great choice if you need high transduction efficiency.

We do not validate shRNA sequences designed as a part of the shRNA cloning services. However, we will send you the shRNA plasmids to test. Once you determine which shRNA construct works best for your experiments, we can package your selected shRNA construct into virus.

By default, expression of our lentivirus ORF cDNA clones are driven by the CMV promoter. C-terminal FLAG and Myc tags are included to allow for easy detection and purification of the overexpressed protein.