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AAV Titers: Where do they come from and what do they mean?

Posted by Leila Haery on Nov 15, 2017 10:00:00 AM

Remember the game show “The $25,000 Pyramid” where one player tries to get the other to guess a category by listing off things that fall into that category? Okay, let’s play! I’ll list the examples and you try to guess the category:

ELISA...
qPCR...
Digital droplet PCR...
DNA dot blot...
Transduction assay...
SDS-PAGE...
Electron microscopy…

Any guesses?

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Topics: Viral Vectors

Adeno Associated Virus (AAV) for Cell and Gene Therapy

Posted by Guest Blogger on Nov 7, 2017 8:59:12 AM

This post was contributed by guest blogger Harshana S De Silva Feelixge.

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Gene therapy technologies hold great promise for improving or potentially curing human diseases that were previously thought to be incurable. Rapid advances in next generation sequencing technologies have allowed scientists to quickly identify underlying genetic causes of some human conditions, opening up new avenues for therapeutics that treat disease at the molecular level. For instance, if a disease is caused by a mutation in a single gene, it can potentially be treated by correcting the mutation or replacing the gene. A notable example is the treatment of Severe Combined Immune Deficiency disease (SCID-XI), also known as bubble boy syndrome. This disease is caused by mutations in the common cytokine receptor gamma chain (c) and is characterized by a lack of immune cell development and function. To date, gene therapy has been used to treat 10 infants with this disease. To do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were transferred back into the infants. Almost all patients have achieved persistent immunological reconstitution with a normally functioning T cell repertoire (1).

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Topics: Viral Vectors

AAVs CREATed for Gene Delivery to the CNS and PNS

Posted by Beth Kenkel on Sep 28, 2017 10:01:35 AM

Adeno-associated viral (AAV) vectors are the most frequently used gene-transfer tools in the study of the brain and spinal cord, which together are known as the central nervous system (CNS). AAVs are popular tools because: 1) their genomes are easy to manipulate, 2) they have long-term expression; and 3) they have limited toxicity. However, a key challenge of using AAVs for neuroscience research is the lack of a method for genetically manipulating neurons throughout the whole brain. Neurons of the peripheral nervous system (PNS), which connect the heart, lung, gut, and other organs to the CNS, are also an important target for gene delivery, especially for the study of pain. While many new capsids (i.e. the part of the virus that determines tropism) have been developed that increase transduction efficiency, none allow for simple and efficient transduction of both the CNS and PNS.That is until the Gradinaru Lab at Caltech stepped up to the challenge.

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Topics: Plasmid Technology, Hot Plasmids, Viral Vectors

AAV Vector Quality Control: Going the Extra Mile with NGS

Posted by Karen Guerin on Sep 12, 2017 9:44:59 AM

Reproducible data are key to science, so scientists are used to repeating experiments to confirm their findings. But no scientist wants to repeat an experiment because of poor reagent quality. To make sure our AAV vectors are of the highest quality, we undertake a rigorous quality control process - read on to learn more!

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Topics: Inside Addgene, Viral Vectors

How Dry Ice Affects Sample pH & How to Avoid It

Posted by Leila Haery on Jun 20, 2017 10:30:00 AM

We’re always looking for ways to improve our shipment processes. After reading a publication describing how short term storage on dry ice can shift sample pH, we wondered whether or not the dry ice we use to keep viruses frozen during shipment was having an impact on the samples. We therefore devised a few experiments to determine if our tubes were permeable to the CO2 released from dry ice, and whether this affected the pH of our viral samples. Read on to learn how aqueous samples might be affected by dry ice, and specifically how dry ice can affect virus from Addgene.

Bottom line: there’s good news and there’s bad news. The bad news is that some of tubes’ o-rings are, in fact, permeable to CO2 at low temperatures (-80°C) and once in the tube, the CO2 can alter the pH of the liquid sample. The good news is that this effect is reversible and the pH shift can be prevented. Keep this information in mind if you’re planning on shipping something on dry ice or if you’re receiving samples on dry ice - it may prevent you from seeing some unexpected results.

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Topics: Viral Vectors

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