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Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing
Published on: January 7, 2019
Generation of recombinant herpes simplex virus ampiicons
1Department of Antiviral Therapy, Schering-Plough Research Institute, Kenilworth, NJ.
Methods in Molecular Medicine
|March 5, 2011
Summary
The herpes simplex virus type 1 (HSV-1) amplicon system enables efficient gene transfer into mammalian cells. This defective viral vector facilitates high-level expression of foreign genes, including Hepatitis C virus (HCV) proteins, for research and potential gene therapy applications.
Area of Science:
- Virology
- Molecular Biology
- Gene Therapy
Background:
- Herpes simplex virus type 1 (HSV-1) amplicon vectors are novel eukaryotic expression tools.
- These vectors contain an HSV-1 origin of replication (ori) and a packaging (pac) signal for genome processing.
Purpose of the Study:
- To describe methods for utilizing the HSV-1 amplicon expression system.
- To demonstrate high-level expression of Hepatitis C virus (HCV) NS3/4A complexes in mammalian cells.
Main Methods:
- Amplicon vectors are amplified into concatemers and packaged into defective HSV-1 viral particles using helper viruses.
- High-titer defective HSV-1 amplicon virus stocks are generated and their titers determined.
- HCV NS3/4A complexes are expressed using the defective amplicon viruses.
Main Results:
- The amplicon system allows for efficient transfer of foreign genes into a wide range of mammalian cells.
- High levels of HCV NS3/4A complexes were produced in mammalian cells.
- The system is suitable for studying HCV protein expression in various cell lines, including human hepatocytes.
Conclusions:
- The HSV-1 amplicon system is a powerful tool for high-level gene expression and gene therapy.
- It offers a valuable platform for investigating HCV protein biology in relevant cellular models.
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