What is an AAV promoter?

The promoter is a major DNA regulatory element in the viral vector genome that determines the level of transgene expression and in which cells the transgene will be expressed. The choice of promoter is therefore an essential aspect of the design of AAV vectors.

How long does AAV expression last in liver?

Importantly, the AAV genome persisted in most tissues, including the liver, for at least one year (Figure 1) and continued to mediate high level expression (8). Transduced viral cDNA persists in most tissues for at least 1 year.

What are AAV therapies?

Simply put, AAV is transformed from a naturally occurring virus into a delivery mechanism for gene therapy. The viral DNA is replaced with new DNA, and it becomes a precisely coded vector and is no longer considered a virus, as most of the viral components have been replaced.

What is AAV and why is it called AAV?

Adeno-associated viruses (AAV) are small viruses that infect humans and some other primate species. They belong to the genus Dependoparvovirus, which in turn belongs to the family Parvoviridae.

How do you infect cells with AAV?

Infecting cells with AAV: Incubate cells with the virus-containing media for 6-12 hours, or as long as you wish. (Optional), you could remove virus-containing media and replace it with fresh, desired media. The appropriate amount of viruses used for infecting cells is critical for the outcome of your experiments.

What cells target AAV?

AAV-2 has a broad tissue tropism infecting diverse organs such as brain, liver, muscle, lung, retina and heart muscle. This makes AAV-2 attractive for in vitro gene transfer into various tissues.

How long does AAV expression last?

showed that an AAV vector would continue to express its transgene for 6–12 months in vivo. Subsequently, expression from an AAV vector in a canine eye persisted unabated for up to 12 years (William Hauswirth, unpublished), and similar results have been reported for muscle and brain transductions.

How long does AAV expression last in mice?

AAV-mediated EGFP expression lasted for at least 1 year. AAV vectors instilled into the pericardial space transduced epicardial myocytes. Arterial gene transfer was studied in mouse carotids (n=26). Both vectors selectively transduced endothelial cells after luminal instillation.

How do you target an AAV?

Strategies for targeted manipulation include combinations of classical genetic tools such as Cre/loxP and Flp/FRT, use of cis-regulatory elements, targeted knock-in transgenic mice, and gene delivery by AAV and other viral vectors.

How does AAV-mediated liver-directed gene therapy work?

AAV-mediated liver-directed gene therapy. The liver is directly or indirectly involved in many essential processes and is affected by numerous inherited diseases. Therefore, many inherited diseases could be effectively treated by targeting the liver using gene transfer approaches.

Where to find liver specific enhancer promoter activity?

1 Gene Therapy Research Unit, Children’s Medical Research Institute and Sydney Children’s Hospitals Network, University of Sydney, Sydney, New South Wales, Australia. 2 Bioinformatics Unit, Children’s Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia.

Is there a gene therapy for liver disease?

Adeno-Associated Virus Gene Therapy for Liver Disease. Abstract. The field of adeno-associated virus (AAV) gene therapy has progressed rapidly over the past decade, with the advent of novel capsid serotype and organ-specific promoters, and an increasing understanding of the immune response to AAV administration.

How is the proteome of the liver related to specificity?

Transcriptome analysis of the liver can be visualized with regard to specificity and distribution of transcribed mRNA molecules (Figure 1). Specificity illustrates the number of genes with elevated or non-elevated expression in the liver compared to other tissues. Elevated expression includes three subcategory types of elevated expression: