What are AAV injections?

Intravenous injections are primarily used for peripheral tissue delivery but with the discovery of novel AAV capsids that can cross the blood brain barrier, intravenous injections can be used to deliver therapeutic rAAV vectors to the CNS of neonatal mice.

Is GFAP specific for astrocytes?

Glial fibrillary acidic protein (GFAP) is an intermediate-filament protein expressed abundantly and almost exclusively in astrocytes of the CNS.

What is the function of astrocytes in the brain?

Astrocytes are the most numerous cell type within the central nervous system (CNS) and perform a variety of tasks, from axon guidance and synaptic support, to the control of the blood brain barrier and blood flow.

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.

How long does an AAV expression take?

Timing will highly depend on the capsid type and on the tissue you’re infecting. Waiting ~2 weeks is a good starting point for many tissues. AAV-mediated gene expression has been reported to be quite stable, lasting for several years in human clinical trials and in dogs (Wonjo et al., 2013).

Where is GFAP found in the brain?

astrocyte cells
GFAP is expressed in the central nervous system in astrocyte cells. It is involved in many important CNS processes, including cell communication and the functioning of the blood brain barrier.

How do astrocytes protect the brain?

Astrocytes are capable of producing a robust antioxidant response to protect themselves and also neurons, through the release of glutathione precursors to neurons. Their role in scar formation allows astrocytes to regulate and contain the immune responses in a manner that controls neuroinflammation.

What are the two functions of astrocytes?

Functions of astrocytes include physical and metabolic support for neurons, detoxification, guidance during migration, regulation of energy metabolism, electrical insulation (for unmyelinated axons), transport of blood-borne material to the neuron, and reaction to injury.

How long does AAV last?

and Xiao et al. 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 is astrocyte-specific gene expression mediated by AAV?

AAV-Mediated Astrocyte-Specific Gene Expression under Human ALDH1L1 Promoter in Mouse Thalamus Adeno-associated virus (AAV)-mediated gene delivery has been proposed to be an essential tool of gene therapy for various brain diseases.

Which is better for astrocytes AAV9 or AAV5?

AAV5 resulted in greater transduction efficiency, transgene expression and astrocyte tropism compared with AAV9 and AAVRec2. In a rodent model of SCI, robust transgene expression by AAV5-GFAP/GfaABC 1 D-dYFP was observed through 12 mm of spinal cord tissue and expression was largely restricted to astrocytes.

Which is the best gene therapy for astrocytes?

Gene delivery towards astrocytes may be beneficial in addressing their role in SCI. Adeno-associated viral (AAV) vector gene therapy, in particular, has emerged as the vector of choice for safe, robust and long-term transgene expression in the CNS.

How are AAV serotypes selected for gene delivery?

AAVDJ8 displayed more tropism in astrocytes compared to AAV9 in the SN region. We conclude that ICV injection results in lasting expression of virally encoded transgene when using AAV vectors and that specific AAV serotypes are required to selectively deliver transgenes of interest to different brain regions in both astrocytes and neurons.