Guest blogger Todd Waldman, Professor at Georgetown University, contributed to this post.
Adeno-associated viruses (AAVs) make fantastic gene delivery vehicles for episomal gene expression and are particularly useful for gene delivery to the nervous system. For many years they have also been used to enhance the efficiency of genome editing. In this post we'll walk through a variety of ways you can use AAVs to improve your genome editing experiments (with and without targeted nucleases).
Topics: Hot Plasmids, Viral Vectors
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).
Topics: Viral Vectors
As the saying goes, “failing to plan is planning to fail.” Preparation is a key step to any experiment and can help prevent future headaches. To help you plan PEI transfections for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires transfecting three plasmids at specific molar ratios. To get these ratios right, you need to do a few calculations. This calculator can do the math for you and it’s simple to make adjustments for the type of tissue culture dish and number of dishes used. Here’s to making AAV packaging a little easier!
Topics: Viral Vectors
We began distributing ready-to-use virus preps through our viral service in late 2016 and requests are still pouring in! While our lentiviral service is going strong, the AAV service has shown incredible growth this year. pAAV-hSyn-DIO-hM4D(Gi)-mCherry was the top requested AAV prep for the 2nd year running, and you can learn more about this useful, DREADD-containing AAV here. But the top requested AAV that became available in 2017 is pmSyn1-EBFP-Cre from Hongkui Zeng’s lab. This AAV has had over 150 orders since coming online!
Topics: Plasmid Technology, Hot Plasmids, Viral Vectors
Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances biological research and discovery. CRISPR is a great example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9 and other genes to mammalian cells. For this reason, the top requested plasmid of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’s lab!
Topics: Plasmid Technology, Hot Plasmids, Viral Vectors
