Pancreas Targeting AAV Production Services

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 used in our vector cloning services. As an outcome, we have a growing base of highly satisfied customers who come back to us time after time for their AAV vector cloning and AAV packaging requirements.

The pancreas is a primary site of origin for a broad variety of diseases, comprising diabetes, pancreatic cancer, and pancreatitis. Pancreatic ductal adenocarcinoma (PDAC) has the terrible prognosis among pancreatic disorders. In the United States, it is the third leading justification of cancer deaths and is projected to become the second leading cause of cancer-related deaths in just over a decade.

A vast range of non-viral and viral gene delivery systems have been exploited to target the pancreas for therapeutic purposes and functional analyses. Non-viral liposomes and nanoparticles have been generally employed to deliver conjugated drugs/chemotherapy or therapeutic genes. However, a main limitation with non-viral delivery procedures is low efficacy, and they need repeated delivery to fulfill therapeutic benefit. Viral vectors have been shown to be more effective to fulfill long-term gene expression with limited off-target effects

Recombinant adenoviruses (rAds) or adeno-associated viruses (rAAVs) are predominately episomal while still attaining efficient in vivo pancreatic delivery. Ad vector-mediated pancreatic gene transfer is transient in behavior because of potent host immune responses and vector-mediated cytotoxicity.  On the other hand, non-integrating AAV vectors are especially favorable in efficiently targeting the whole pancreas, the likely to target specific cell types in the pancreas and pancreatic neoplasms and do not stimulate humoral responses or changes in pancreatic functions.

Among various AAV serotypes tested, AAV8 and AAV9 are well-suited for systemic delivery, and AAV6 have been indicated to transduce the normal pancreas very efficiently via retrograde pancreatic ductal delivery. Accumulating evidence documents the aim of AAV to deliver therapeutic molecules/genes to neoplasms, comprising pancreatic tumours and tissues undergoing rapid degeneration and regeneration.

Production Process

Suitable serotypes for Lung tissue

AAV8

Subcloning (Optional)

We begin by subcloning the gene of interest (GOI), shRNA or gRNA into an associated pAAV cis-plasmid.

Large-scale Production (Optional)

Large-scale preparation of the pAAV cis-plasmid and complimentary plasmids utilizing Qiagen Endo-free Mega Prep kits.

Large scale Transfection

Large-scale transfection of engaged plasmids into 40x15cm plates of HEK293 cells.

Purification

Harvest the AAV production cells and purify the AAVs through a sequence of CsCl centrifugations.

Titer Determination 

Find the titer of the viral stock (in genome copy number per ml, or GC/ml) through quantitative real-time PCR.

Workflow

Genome-wide AAV Products

Human

•Over-Expression Products

•shRNA Products

•miRNA Products

Mouse

•Over-Expression Products

•shRNA Products

•miRNA Products

Rat

•Over-Expression Products

•shRNA Products

•miRNA 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 

References

1.Dimastromatteo J., Brentnall T., Kelly K.A. Imaging in pancreatic disease. Nat. Rev. Gastroenterol. Hepatol. 2017;14:97–109.

2.Von Hoff D.D., Ervin T., Arena F.P., Chiorean E.G., Infante J., Moore M., Seay T., Tjulandin S.A., Ma W.W., Saleh M.N. Increased survival in pancreatic cancer with nab-paclitaxel plus gemcitabine. N. Engl. J. Med. 2013;369:1691–1703.

3.Ryan D.P., Hong T.S., Bardeesy N. Pancreatic adenocarcinoma. N. Engl. J. Med. 2014;371:2140–2141.