An Engineered Virus Library as a Resource for the Spectrum-wide Exploration of Virus and Vector Diversity

Publication date: 23 May 2017
Source:Cell Reports, Volume 19, Issue 8
Author(s): Wenli Zhang, Jun Fu, Jing Liu, Hailong Wang, Maren Schiwon, Sebastian Janz, Lukas Schaffarczyk, Lukas von der Goltz, Eric Ehrke-Schulz, Johannes Dörner, Manish Solanki, Philip Boehme, Thorsten Bergmann, Andre Lieber, Chris Lauber, Andreas Dahl, Andreas Petzold, Youming Zhang, A. Francis Stewart, Anja Ehrhardt
Adenoviruses (Ads) are large human-pathogenic double-stranded DNA (dsDNA) viruses presenting an enormous natural diversity associated with a broad variety of diseases. However, only a small fraction of adenoviruses has been explored in basic virology and biomedical research, highlighting the need to develop robust and adaptable methodologies and resources. We developed a method for high-throughput direct cloning and engineering of adenoviral genomes from different sources utilizing advanced linear-linear homologous recombination (LLHR) and linear-circular homologous recombination (LCHR). We describe 34 cloned adenoviral genomes originating from clinical samples, which were characterized by next-generation sequencing (NGS). We anticipate that this recombineering strategy and the engineered adenovirus library will provide an approach to study basic and clinical virology. High-throughput screening (HTS) of the reporter-tagged Ad library in a panel of cell lines including osteosarcoma disease-specific cell lines revealed alternative virus types with enhanced transduction and oncolysis efficiencies. This highlights the usefulness of this resource.

Graphical abstract

Teaser

Zhang et al. develop a direct cloning strategy to incorporate viral genomes into an easy-to-handle plasmid, thereby generating a library consisting of cloned adenovirus plasmids and reporter-tagged vectors. This library, together with the homologous recombineering technique, offers a resource to study pathogenesis and to establish therapeutic applications.


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