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

Cell Rep. 2017 May 23;19(8):1698-1709. doi: 10.1016/j.celrep.2017.05.008.

Abstract

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.

Keywords: Red/ET homologous recombination; adenovirus; engineered adenovirus library; large double-stranded DNA viruses; natural virus diversity; resource; therapy; translation; vector.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Base Sequence
  • Cloning, Molecular
  • Gene Library*
  • Genes, Reporter
  • Genetic Engineering*
  • Genetic Vectors / metabolism*
  • Genome, Viral
  • High-Throughput Screening Assays
  • Humans