Adenoviral FAQs

Adeno-X Adenoviral System 3 from Takara Bio is the most advanced commercially available adenoviral gene delivery system. It is a single-vector-based system that allows you to clone a gene of interest (GOI) or custom-made expression cassette directly into the adenovirus backbone in a single step, using In-Fusion technology.

Vector cloning/packaging capacities vary, depending on the specific adenoviral vector backbone:

Our adenoviral vectors carry deletions in the E1 and E3 regions of the adenovirus genome.

Our adenoviral vectors are derived from human adenovirus type 5.

Yes. We offer a third-generation Tet-On 3G inducible expression system in a single adenoviral vector format, the Adeno-X Adenoviral System 3 (Tet-On 3G Inducible).

Takara Bio has validated the Adeno-X purification kits for purification of Adeno-X viruses based on type 5 human adenovirus.

No. The Adeno-X Maxi/Mega purification kits only purify virus from cell pellets, which accumulate most of the amplified virus, provided that the cells are harvested prior to complete cell lysis (Figure 1). This protocol greatly simplifies adenoviral purification.

Figure 1. Time course of adenoviral yields from cell pellets and supernatants. HEK 293 cells were transduced with an Adeno-X construct expressing DsRed-Express and incubated for 2–4 days. On Days 2–4, cells were harvested, lysed, and centrifuged—and each pellet (containing the cell’s contents) and supernatant (containing the medium the cells were growing in) was used to infect fresh HEK 293 cells to measure titer. Most of the virus was found in the cell pellets, especially by Day 4, with only 2% in the supernatant.

Yes. Adenovirus purified with Adeno-X purification kits can be used for in vivo experiments after desalting by column chromatography or dialysis. We recommend using the Adeno-X Mega Purification Kit (Cat. # 631032) to purify adenovirus for in vivo experiments.

The Adeno-X Rapid Titer Kit is compatible with human adenoviruses (types 1, 2, 5, and 6). Please note that we have validated the Adeno-X Rapid Titer Kit for titration of Adeno-X viruses, based on type 5 human adenovirus.

MOI, or multiplicity of infection, is the number of infectious viral units per cell at the time of infection.

IFU, or infectious units, determines the virus’ ability to infect cells and express protein. You can determine the infectious viral titer (IFU/ml) by measuring the expression of a reporter or a specific protein by flow cytometry, immunostaining, etc.

PFU, or plaque forming units, measures the ability of the virus to infect permissive cells and replicate, forming plaques (holes that represent lysed cells) in the cell monolayer. The titer in PFU/ml is determined by a plaque assay.