Organelle Probe Adenovirus

  • Product Overview
  • Manuals & MSDS
  • FAQs
  • Citations and references
  • Recommended products

Introduction to Organelle Probe Adenovirus

Recombinant adenovirus (Adenovirus), a replication-defective adenoviral vector system, is widely used for gene delivery in most cell types. 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. Adenovirus 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.

Properties

Adenovirus
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 > 5ug each

Advantages

Broad range of host.Adenovirus has the ability to infect dividing, quiescent cells, stem cells, and primary cells, allowing genetic materials to be delivered to a highly diverse range of cell types and tissues.

Applications and Figures

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Figure 1
Transduction results of various Adenovirus vectors. Selected results of Adenovirus transduction in the adult rat cardiac cells (A), eyes (B), neuron cells (C).

Technical Documents

1. For detailed protocol about how to package and purify Adenovirus, please see the pdf downloadAdenovirus User Manual.

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.

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Reference

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