Oncolytic herpes simplex virus engineering and preparation
Abstract
Herpes simplex virus-1 (HSV-1) is an enveloped, double-stranded DNA virus that has been used with modification as an oncolytic virus against a number of tumor types. Modifications that make HSV-1 replication--conditional, i.e., selectively divide in replicating cells make it fulfill a prerequisite criteria for oncolytic viruses. Other appealing features of HSV-1 as an oncolytic virus include its large, modifiable genome; its sensitivity to antiviral agents, such as ganciclovir; and its lack of host cell integration. Here, we review the methods of HSV-1 engineering, through traditional recombination techniques as well as through bacterial artificial chromosome (BAC) technology. We then describe protocols for titering, amplification, and purification of engineered HSV-1-derived oncolytic viruses.
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Prior methods for HSV1 genome editing require amplification and purification to obtain viral genomic DNA, then co-transfection with homologous DNA sequences into cells followed by two-step counter-selection and multiple plaque isolation or insertion into a BAC system and several in vitro molecular biological procedures.
“The genomic DNA is then co-transfected with homologous DNA sequences into cells followed by two-step counter-selection and multiple plaque isolation [30] or by inserting the viral genome into a BAC system and performing several in vitro molecular biological procedures [31].”
From High-efficiency targeted editing of large viral genomes by RNA-guided nucleases (2014) · cites this paper as [31]
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- Authors
- Agarwalla, P. K., & Aghi, M. K.
- Affiliations
- Harvard University (US)
- University of California, San Francisco (US)
- Corresponding author
- Aghi, Manish K.
- ORCID
- Aghi, Manish K.
- Venue
- Methods in molecular biology
- Publisher
- Humana Press
- Published
- 2011-08-23
- Volume
- 797
- Pages
- 1-19
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- OpenAlex
- W2406631797
- Cited by
- 37
- References
- 12
- Citations by year
- 2026: 1 · 2024: 3 · 2023: 4 · 2022: 3 · 2021: 5 · 2020: 2 · 2019: 2 · 2018: 5 · 2017: 3 · 2016: 3 · 2014: 1 · 2013: 3 · 2012: 1
- FWCI
- 4.90
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Doris Duke Charitable Foundation
- American Brain Tumor Association
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Health Sciences › Medicine › Epidemiology › Herpesvirus Infections and Treatments
- Health Sciences › Medicine › Oncology › Viral-associated cancers and disorders
- Keywords
- oncolytic herpes simplex virus, herpes simplex virus type 1, conditional replication, bacterial artificial chromosome, viral recombination, ganciclovir sensitivity, viral replication, virus purification, antiviral resistance
- MeSH terms
- Antiviral Agents, Genetic Engineering, Genetic Vectors, Humans, Recombination, Genetic, Virus Replication, Ganciclovir, Herpesvirus 1, Human, Chromosomes, Artificial, Bacterial, Oncolytic Virotherapy, Oncolytic Viruses
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