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Construction of Oncolytic Adenovirus Vectors

Oncolytic adenoviruses (a.k.a Conditionally-Replicative Adenoviruses: CRAds) are  viruses that can selectively kill cancer cells while leaving healthy cells intact.  They are generally constructed by placing viral genes involved in virus replication (e.g. the E1 region) under the control of tumor-specific elements.  The killer effect of these oncolytic viruses can be potentiated by expressing "therapeutic" genes such as pro-apoptotic genes,  from the same vector, thereby generating so-called "armed CRAds".  These viruses have the potential to cure cancers that are resistant to current treatments such as chemotherapy and radiation.

O.D.260 Inc has more than 15 years experience in constructing oncolytic adenoviruses for clients around the world.  We have the tools to construct your recombinant CRAd easily and at low cost.

Applications

  • Oncolytic adenovirus vectors with the E1 region under the control of a tumor-specific promoter
  • Oncolytic adenovirus vectors with the E1 region under the control of a tumor-specific promoter and with a transgene expressed in the E3 region ("Armed" oncolytic vectors)

Features/options

  • Cloning systems: AdenoQuick1.0, AdenoQuick2.0
  • Backbone: Ad5
  • Packaging signal: complete (all seven "A" repeats) or partial (repeats A1-A5)
  • E1 region:  WT or promoter region deleted, with E1a TATA box present (Δ bp 354-462) or deleted (Δ bp 354-480)
  • E3 region: WT, or deleted (1.6 or 2.7 kb) with multiple cloning site
  • E4 region: WT, or deleted (1.2 or 2.8 kb)
  • Fiber: WT Ad5, hybrid Ad5/35, Ad5/3
  • Maximum cargo capacity: 10.4 kb

The service includes

  • Design of a cloning strategy
  • Construction of a cosmid containing the entire genome of your oncolytic adenovirus
  • If necessary, verification of the DNA sequence
  • Transfection-grade plasmid DNA purification
  • Virus rescue by transfecting the linearized cosmid DNA into helper cells
  • Virus clone isolation
  • Confirmation of virus identity at the DNA level

You can customize your request: for instance, you can decide to have us construct the adenovirus cosmid, and perform the virus rescue yourself.

Deliverables

  • The crude virus lysates of three clones of your recombinant virus (1 mL per clone)
  • A complete report describing the work performed and demonstrating the identity of the virus clones we are sending to you

Turn-around time

  • About a month, depending on how many cloning steps in E. coli are necessary

Pricing

  • Inquire

Important note

  • The use of adenovirus as a genetic vector requires adequate containment equipment and practices. Biosafety Level 2 (BL2) is appropriate for most customers; however, for some, use of BL3 or higher level containment facility may be required. O.D.260 Inc. will not carry out projects that require BL3 (or above) containment.