Knock-out of a gene of interest is a useful gene-editing strategy to validate a gene function, to develop a synthetic lethality model or directly mimic a cancer cell in drug screening. Unfortunately, non-dividing cells and hard-to-transfect cells are a major challenge for such genome-editing. However, there is an ideal solution that deserves to be more popular, the use of adenovirus.

What makes adenovirus so good for KO?

First of all, adenovirus is one of the best viral vectors for cell culture infection in terms of transduction efficacy, with nearly 100% efficiency even on hard-to-transfect cells. The picture at the top of this post illustrates this with a HepG2 cell line and Ad-GFP, an adenovirus expressing GFP under CMV promoter (image legend: Ad-GFP transduction into HepG2 cells).

And that’s not all. Adenovirus cannot be inserted in the genome. Thus, there is no risk of genotoxicity or non-specific KO. It’s safer.

And last but not least, the expression is transient. And such a temporary expression of CAS9 leads to a lower risk of off-target.

Any drawbacks related to adenovirus-based KO then?

Adenovirus expression is so high that it leads to inflammation. However, high inflammation happens only when the adenovirus is injected into animals. For in vitro cell culture infection, adenovirus is perfect.

How to benefit from adenovirus-based KO?

In Europe, SignaGene laboratories, via tebu-bio, offers gRNA and CAS9 expressing adenovirus. The structure of the AdV is like “ITR-CMV-Cas9-polyA-U6-gRNA-polyT-ITR”.

Researchers can provide their preferred sgRNA sequence or we can design it to be of further assistance.

The final deliverable is super infectious gRNA AdV vector 1000 uL at >5×10^10 PFU/mL. That concentration corresponds to high grade quality.

Do you want to develop a monoclonal cell line? High transduction efficiency combined with the smart aliquoter make it a piece of cake.

In conclusion, to get your gene editing project started, just mention your target gene below – we’ll be pleased to get back to you!

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    Written by Dimitri Szymczak, PhD
    Dimitri Szymczak is a Technical Support Specialist and Product Manager at tebu-bio, and a fan of capoeira in his spare time.