Understanding the biology behind brain development, learning, and behavior poses many challenges in neuroscience with many brain regions, neuronal networks, and specific neuronal populations to consider. To tease apart how different neurons impact a certain output, neuroscientists have begun to use more precise genetic tools: activators, inhibitors, sensors, recombinases, and more. With the help of adeno-associated viral vectors (AAV), neuroscientists can combine classical genetic tools with the manipulation of specific neuronal subtypes and determine how these changes affect the phenotype. Viral vectors and genetic approaches are continuously refined and developed so there are many combinations to choose from for your experiments!

Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production, assembled AAV vector particles accumulate inside the producer cells and purification protocols often describe a process that begins with cell lysis to release AAV particles with subsequent purification steps. It’s also possible for AAV vectors to be spontaneously released from the producer cells into the supernatant during the production phase. However, this has not been well characterized as it is not a common event.

By now you have probably heard about Addgene’s viral service but did you know that Addgene also performs independent research? Since launching the viral service in 2016 Addgene’s research team has been actively investigating ways to improve viral vector production. Whether redesigning viral packaging plasmids, investigating different viral production strategies or developing novel methods for data analysis, our team’s goal is to provide scientists with new tools and protocols that they can easily incorporate into their own labs. This week we are pleased to announce the submission of the results of our first study which can be accessed here at bioRxiv.

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.