Also be very careful about the promoter driving your gene of interest in the virus : CMV often gets down-regulated in neurons, for instance. You may want to try EF1alpha or other promoters.

I have heard good things about the AAV system from Cell Biolabs. They offer the shuffled capsid version called AAV-DJ and AAV-DJ/8 (more like AAV8 in terms of modification of the heparin binding domain). These shuffled capsid proteins give infection of a broader range of cell types. In addition they offer a nice antibody matrix based column purification system called Virabind that can be used to purify and concentrate the virus. The production is done by triple transfection of AAV-293 cells, also from Cell Biolabs.

In my own hands I have been using AAV293 cells, AAVrep2-cap8 plasmid, and pAd-deltaF6 plasmid. The DNA was obtained by MTA and purchase from UPenn. The ITR containing vector for cloning genes of interest was obtained from Addgene, and there are tons of available vectors to choose from depending on your desired backbone and features. Thus, you can piece your system together fairly cheaply without going for an out of the box commercial system.

Hi, Jonathan,

I am trying to purify the AAV DJ/8. Do you know whether there is kit for the purification?

Fangqiang

Hi Fangqiang, I have been told that there is a commercial kit that can purify serotype 8 (or equivalent DJ/8). The company is called Genicity

http://genicity.com/index.php?dispatch=categories.view&category_id=454

However, I do not have direct experience with this product. I hear you may be able to get a free sample to test.

Jonathan T Ting,

Can I ask you about the AAV2-cap8 plasmid that you use? Where do you get it from?

Thank you

someone asked about the Delta F6 helper sequence.

I hope it serves you.