Recombinant Adenovirus Rescue in Mammalian Cells
Adenoviral (Ad) vectors are a versatile tool in the investigations of gene expression and regulation as well as gene therapy. Over the years, many approaches have been developed for the generation of recombinant adenoviruses. As a pioneer service provider for gene therapy development, Creative Biolabs is capable of providing high-quality recombinant Ad vectors construction services to promote the development of global customer's projects. Here, we are happy to introduce our recombinant Ad rescue in mammalian cells.
Homologous Recombinant in Mammalian Cells
Methods for the generation of recombinant adenoviruses can be divided into two basic categories, namely direct plasmid construction of recombinant adenoviral genome and indirect construction. The former involves the ligation of the adenoviral genome with the DNA fragments of interest, and the latter involves homologous recombination in mammalian cells or in Escherichia coli. Homologous recombination in mammalian cells is completed through a 'shuttle' recombined with a 'backbone' plasmid. Typically, as shuttle vector contains the 5' end of the adenoviral genome wherein the E1 and the interested gene are located. And the backbone vector is the primary carrier of the majority of the viral genome but importantly lacks essential genes for viral propagation in naturally occurring cells. After recombinant, a single DNA molecule which encodes all genes required for viral replication and production exclusively in 'packaging cell lines' but not in naturally occurring cells is generated.
Figure 1. Schematic representation of the DNA vector-mediated homologous recombination in mammalian cells. (Nafissi, 2015)
Homologous Recombinant Ad Construction Service Provided by Creative Biolabs
Despite construction Ad vector without repeated rounds of plaque purification, the generation of recombinant Ad is still limited by several factors, including the low efficiency and difficulty in the screening of homologous recombination, the need for time-consuming plaque purification, and the frequent contamination by wild-type adenoviruses. Considering the aforementioned drawbacks, we have now developed robust and scalable platforms to generate recombinant adenoviruses based on previously reported cloning system. Our modified methods have made it possible to generate large quantities of recombinant adenoviruses in a timely and predictable manner.
Features of Our Services
- Large packaging capacity
- High efficiency of expression that can often be applied in research within 24 hours
- High purity for safe use in vivo
- Strong immunogenicity
With world-class technology of recombinant vectors construction, Creative Biolabs provides customized Ad vector production with robust quality control. Our professional scientists are happy to tailor the most reasonable scheme to satisfy different experimental needs. If you need any help in recombinant Ad vector design and construction, please feel free to contact us for more details.
Reference
- Nafissi, N.; Foldvari, M. (2015). Neuroprotective therapies in glaucoma: II. Genetic nanotechnology tools. Frontiers in neuroscience. 9: p.355.
