Heart Targeting AAV Production Services

Seattle Genova offers outstanding quality AAV packaging services to benefit your AAV-based gene therapy experiments. We have enhanced a series of proprietary technologies and reagents that have largely improved recombinant AAV production protocols in terms of titer, purity, potency and consistency, particularly for the AAV vector systems utilized in our vector cloning services. As a consequence, 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.

In the field of cardiovascular disease (CVD), compared with non-viral vectors, lentiviruses, poxviruses, and adenovirus vectors, AAV possesses various advantages, containing high security, low immunogenicity, sustainable and stable exogenous gene expression etc., which makes AAV one of the greatly promising candidates for the treatment of several genetic disorders and hereditary diseases.

Suitable Serotypes for Heart tissue

AAV1, AAV8, AAV9

Application of AAV vectors to gene therapy of cardiovascular disorder

Recombinant AAV and heart failure

HF, the leading reason of death from cardiovascular diseases, is a serious progressive disease that harms the heart and its purpose and irreversibly worsens without any therapeutic intervention. 

Appreciating the pathological mechanisms and natural record of HF is the source for accomplishing needed treatment outcomes. According to a vast number of randomized controlled trials interventions that increase left ventricular systolic function are the major treatment modalities.

Recombinant AAV and cardiomyopathy

Cardiomyopathy is a heterogeneous disorder that results in systolic and diastolic dysfunction of the heart due to pathological modifications to the myocardium.

Repairing and improving the systolic and diastolic objectives of the heart is a long-term effective technique against cardiomyopathy, and multiple gene therapies have been utilized to enhance cardiac capacity, including cardiac regeneration, enhancing skeletal muscle function, and lessening cardiac hypertrophy. The recent findings show that rAAV9 reversed the impacts of HCM resulting in the D166V mutation by delivering S15D-RLC into the hearts of humanized transgenic HCM-D166V mice, considerably enhancing intact cardiac function.

Production Process

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.Roth GA, Mensah GA, Fuster V. The global burden of cardiovascular diseases and risks: a compass for global action. Journal of the American College of Cardiology. (2020) 76:2980–1. 

2.Nabel Elizabeth G, Plautz G, Boyce Frederick M, Stanley James C, Nabel Gary J. Recombinant Gene Expression in vivo Within Endothelial Cells of the Arterial Wall. Science. (1989) 244:1342–4. 

3.Friedmann T, Roblin R. Gene therapy for human genetic disease? Science. (1972) 175:949–55. 

4.Li C, Samulski JR. Engineering adeno-associated virus vectors for gene therapy. Nat Rev Genet. (2020) 21:255–72.