Adenovirus Production: How to Clone, Package, and Harvest Adenovirus for Your Seed Stocks

Adenoviruses are non-integrating and can deliver large transgenes (up to 6 kb) to a broad range of host cells (both dividing and non-dividing) with high efficiency, providing excellent yields of recombinant protein. In this video, we'll explain how to package second generation adenoviruses and generate high titer seed stocks!
Here are some helpful time stamps for our adenovirus packaging video:
1:50 How to clone your gene of interest into an adenovirus expression system. We recommend abm's Adeno-1 to Adeno-8 cloning kits to easily speed up this process! www.abmgood.com/Adenovirus-Ex...
3:07 How to amplify and linearize your recombinant adenovector for use in the transfection step.
3:41 Preparing your HEK293 cells for transfection (a great adenovirus packaging cell line).
4:21 How to prepare your transfection solutions. We recommend abm's DNAfectin™ Plus transfection reagent (Cat. No. G2500). www.abmgood.com/dnafectin-plu...
5:45 How to transfect your HEK293 cells.
6:15 Checking your cells for successful transfection
6:32 How to make your adenovirus seed stock and amplify it to achieve a higher titer!
7:00 How to harvest and store your adenoviruses.
Learn more about the recombinant adenovirus system at our learning resources center: info.abmgood.com/adenovirus-s...
Browse our collection of ready-to-use adenoviruses for every human, mouse, and rat gene: www.abmgood.com/Adenovirus.html
We also offer ready-to-use CRISPR knockout adenoviruses targeting every human, mouse, or rat gene: www.abmgood.com/CRISPR-Knocko...
Need a custom recombinant adenovirus? We can make it for you! Simply tell us your project details and we'll do the gene synthesis, cloning, and adenoviral packaging for you: www.abmgood.com/Custom-Recomb...
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Here are some helpful time stamps for our adenovirus packaging video:
1:50 How to clone your gene of interest into an adenovirus expression system.
3:07 How to amplify and linearize your recombinant adenovector for use in the transfection step.
3:41 Preparing your HEK293 cells for transfection
4:21 How to prepare your transfection solutions.
5:45 How to transfect your HEK293 cells.
6:15 Checking your cells for successful transfection
6:32 How to make your adenovirus seed stock and amplify it to achieve a higher titer!
7:00 How to harvest and store your adenoviruses.

[@samiurrahmansefat1336]

Why do we linearize adenoviral vector before transfection? The obvious answer might be that it is tough to deliver a large plasmid into cells. Are there any additional reasons?

[@maxbailey9164]

Think you’ve hit the nail on the head. From what I understand, transfection is relatively poorly understood, but I’d imagine it’s similar to the linearisation of a plasmid before being run on a gel