Seattle Genova offers outstanding quality AAV packaging services to benefit your AAV-based gene therapy experiments. We have improved a series of proprietary technologies and reagents that have greatly improved recombinant AAV production protocols in terms of titer, purity, potency and consistency, particularly for the AAV vector systems employed in our vector cloning services. As an outcome, we have a growing base of highly satisfied clients who come back to us time after time for their AAV vector cloning and AAV packaging requirements.
Enhanced vascular permeability in the lungs can direct to pulmonary edema, impaired gas exchange, and eventually respiratory failure. While oxygen delivery, mechanical ventilation, and pressure-reducing medications enable alleviate these symptoms, they do not deal with the underlying disease.
Cell-type Specific Targeting Strategy
The ability to target specific cell types within the lungs has been verify to be extremely helpful for both illness treatment and for prevention and/or diagnosis. Specific to lung, much of the cell-type specific targeting work could be achieved by targeting to alveolar macrophage (AM) and lung cancer cells.
Gene therapy is being investigated for a range of severe lung disorders, such as cystic fibrosis and emphysema. Recombinant adeno-associated virus (rAAV) is a well-established, safe, viral vector for gene delivery with numerous commonly occurring and artificial serotypes accessible displaying alternate cell, tissue, and species-specific tropisms.
The identification of AAV serotypes for gene delivery to the human lung has concentrated primarily on the transduction of the human airway epithelium. The lung parenchyma, however, is the target for treating genetic disorders such as surfactant deficiencies and interstitial lung disease; in particular, alveolar type II (ATII) pneumocytes, which express proteins important for surfactant function.
Adeno-associated virus (AAV)-based vectors are assessed safe and effective, they have been utilized in several clinical trials and one of them has been even approved as first commercial gene therapy drug in the European Union.
Suitable serotypes for Lung tissue
AAV4, AAV5, AAV6,AAV9
We begin by subcloning the gene of interest (GOI), shRNA or gRNA into an associated pAAV cis-plasmid.
2.Large-scale Production (Optional)
Large-scale preparation of the pAAV cis-plasmid and complimentary plasmids utilizing Qiagen Endo-free Mega Prep kits.
3.Large scale Transfection
Large-scale transfection of engaged plasmids into 40x15cm plates of HEK293 cells.
Harvest the AAV production cells and purify the AAVs through a sequence of CsCl centrifugations.
Find the titer of the viral stock (in genome copy number per ml, or GC/ml) through quantitative real-time PCR.
Genome-wide AAV Products
Key Features and Benefits
√Ready to utilize for in vitro and in vivo applications
√High titer: improved transduction efficiency
√Transduction of dividing and non-dividing cells
√Expert Technical Support
√Quality and user friendly
1.Li C, Samulski RJ. Engineering adeno-associated virus vectors for gene therapy. Nat Rev Genet. 2020;21(4):255–72.
2.Moss RB, Milla C, Colombo J, Accurso F, Zeitlin PL, Clancy JP, et al. Repeated aerosolized AAV-CFTR for treatment of cystic fibrosis: a randomized placebo-controlled phase 2B trial. Hum Gene Ther. 2007;18(8):726–32.
3.Yan Z, Lei-Butters DC, Liu X, Zhang Y, Zhang L, Luo M, et al. Unique biologic properties of recombinant AAV1 transduction in polarized human airway epithelia. J Biol Chem. 2006;281(40):29684–92.
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