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

Paper ID Volume ID Publish Year Pages File Format Full-Text
23993 43488 2012 6 PDF Available
Title
A rapid generation of adenovirus vector with a genetic modification in hexon protein
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.

► We developed a novel system for rapid generation of hexon-modified adenoviral vector. ► This vector system is featured by one step ligation and white/blue color screening. ► Adenoviral vector with Tat PTD modification in hexon protein was easily generated. ► Tat PTD modification in hexon markedly improves the Ad transduction in vitro. ► The novel system would be of substantial benefit for cancer gene therapy.

Keywords
Directional cloning; Adenoviral vector; Gene therapy; Quick screening; Hexon modification
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A rapid generation of adenovirus vector with a genetic modification in hexon protein
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 157, Issue 3, 10 February 2012, Pages 373–378
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us