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So you have this awesome experiment you want to do, but it requires some AAV. You’ve never worked with AAV before, but you aren’t going to let that stop you. Where do you start? Turns out like all good experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV: 

1. the packaging plasmid which contains the structural and packaging genes,
2. the helper plasmid which contains the proteins needed for the virus to replicate, 
3. and the transfer plasmid which contains the viral genome.

In today’s blog post, we’ll focus on the AAV transfer plasmid and take a look at each of its parts.

In order to understand the complex wiring of the brain and the underpinnings of neurological disease, neuroscientists need to be able to probe cells and circuits without disrupting normal brain function. Chemogenetics has become a popular tool in the field as it provides a noninvasive method for manipulating cell activity, especially neuronal firing.

My lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, I had never worked with AAV, so naturally I had to package some virus for my first experiment. It was a bit intimidating, but I had my lab’s protocols and some great co-workers to help me out. Even with these tools, I found myself writing AAV production tricks into the margins of my protocol. While these tips weren’t critical to the experiment, they definitely made my life easier!  In this post, I’ll share some AAV production, purification, and titration tips, while also summarizing the basic steps and analyses needed for packaging AAV.

While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges. Whether choosing a system that is the best fit for the experiment, trying to produce virus of a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find themselves faced with a roadblock. In this post, we will provide an overview of some of the common challenges associated with producing and using lentivirus and offer some tips and tricks for overcoming these hurdles.

Many of us take comfort in the fact that it’s often not quantity, but quality that really matters. Well, it turns out this isn’t the case for using AAV. When it comes to infecting cells, titer, the amount of virus used, really does matter. (*psst*, quality definitely also matters).

Check out this post for a refresher on AAV titers