Adenovirus Specific for Suspension Cells

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Introduction to Adenovirus Specific for Suspension Cells

The adenoviral vectors provided by Genemedi are based on human adenovirus type 5 (Ad5), which is replication-incompetent (-E1/-E3) and can’t be integrated into host genome, guaranteeing the security for subsequent operations. Adv packaging by Genemedi shows almost 100% gene delivery in most cell types in the recombinant protein expression system both in vivo and in vitro. Recombinant protein expression of interest is detectable from 24 hours post infection. Furthermore, Genemedi has also developed adenovirus specific for suspension cells, which is modified according to the characteristics of suspension cell membrane. Besides, the virus infection system has also been optimized to obviously increase the infectivity specific for suspension cells. Nevertheless, this virus can also be used to transduce adherent cells, for its infectivity to adherent cells has not changed.

Properties

Adenovirus Virus
Quantity/Unit Vials
Form Frozen form
Sipping and Storage Guidelines Shipped by dry ice, stored at -80 ° C, effective for 1 year. Avoid repeatedly freezing and thawing
Titer > 1*10^10 PFU/ml

Advantages

1. Specific for suspension cells.

2. Extremely high infection efficiency. Adenovirus specific for suspension cells in Genemedi can infect many suspension cells, with almost 100% infection efficiency for Jurat and K562. As for primary cells derived from blood, such as CD4+ T cells, the infection efficiency will be more than 60%.

3. Great packaging capacity, up to 8kb.

4. Without integration. No alteration to host genome.

5. High titer. Up to 1011 pfu/ml.

Applications and Figures

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Figure 1
Transduction results of Adv specific to suspension cells in Jurkat, K562, HL-60, L1210 with fluorescence microscopy at 72 h post infection.

Quality control description

Our optimized custom adenoviral vector production and strict quality control systems provide customers a high titer of functional recombinant adenoviral vectors. Viral titers are determined by TCID50 method, which is the most accurate way to measure the titer of adenovirus.

Technical Documents

1. For detailed protocol about how to package and purify Adv, please see the pdf downloadAdenovirus User Manual on the Genemedi website.

Frequently Asked Questions(FAQs)

1. Why does adenovirus have a relatively higher immunogenecity compared with rAAV?
Answer
The adenovirus without E1/E3 can express all of the other genes in the viral backbone and hence induces immunogenic responses, while rAAV does not have any of the AAV genes, thus no immunogenicity from viral protein.

2. What is the role of the E1 Gene in adenoviruses?
Answer
Simply, the E1 gene products are early proteins that are transcribed in the early transcribed regions and required for proceeding subsequent steps in viral replication. The E1 gene contains E1A and E1B, involved in the replication of adenovirus. E1A is critical to start viral replication by promoting transcription from rep gene promoters, P5 and P19, and facilitate viral replication by activating the early adenovirus promoters.

3. How to store adenovirus?
Answer
It would be better to store adenovirus in PBS at -80oC. Sucrose or DMSO may help to stabilize the vector.

4. How can you tell if your vector is lentiviral, retroviral, or adenoviral?
Answer
You should blast your vector sequence and see if there’re sequences of reverse transcriptase and integrase (gene names: gag and pol), which are for lentiviral/retroviral vectors, but not for adenoviral. For another, if your plasmid is around 30-35kb in size, it's certainly adenoviral.

5. Is adenovirus a useful tool to study primary macrophage functions?
Answer
In RAW264.7 and PM cells, adenovirus works very well, and it seems that IL1β expression is increased slightly after adenovirus transfection compared with negative control. While for the BMDM, adenovirus does not work, it may be better to use lentivirus instead, which gives a pretty good transduction efficacy and less inflammatory response.

6. Is it possible to infect a tissue preparation with lentivirus and afterwards with adenovirus and getting high efficiency in transduction?
Answer
It is definitively possible to perform sequential transductions/infections. Polybreen, protamine sulfate or other transduction enhancing reagents are recommended to enhance viral particle infectivity.

7. How to perform adenovirus transduction in vitro?
Answer
First, amplify to get enough target cells, then infect the target cells with adenovirus. Cell functional assay can be conducted after expression of the target gene. Generally, infected cells will not be amplified for culture.

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Reference

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