A rapid generation of adenovirus vector with a genetic modification in hexon protein

Bingyan Di, Qinwen Mao, Junli Zhao, Xing Li, Dongyang Wang, Haibin Xia*

*Corresponding author for this work

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

The generation of hexon-modified adenovirus vector has proven difficult. In this paper, we developed a novel method for rapid generation of hexon-modified adenoviral vector via one step ligation in vitro followed by quick white/blue color screening. The new system has the following features. First, eGFP expression driven by the CMV promoter in E1 region functions as a reporter to evaluate the tropism of hexon-modified adenovirus in vitro. Second, it has two unique restriction enzyme sites with sticky ends located in the hexon HVR5 region. Third, a lacZ expression cassette under the control of plac promoter is placed between the two restriction enzyme sites, which allows recombinants to be selected using blue/white screening. To prove the principle of the method, genetically modified adenoviruses were successfully produced by insertion of NGR, RGD or Tat PTD peptide into hexon HVR5. Furthermore, the transduction efficiency of the Tat PTD modified virus was shown to be a significant enhancement in A172 and CHO-K1 cells. In conclusion, the novel system makes the production of truly retargeted vectors more promising, which would be of substantial benefit for cancer gene therapy.

Original languageEnglish (US)
Pages (from-to)373-378
Number of pages6
JournalJournal of Biotechnology
Volume157
Issue number3
DOIs
StatePublished - Feb 10 2012

Keywords

  • Adenoviral vector
  • Directional cloning
  • Gene therapy
  • Hexon modification
  • Quick screening

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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