ArticlePloS one2011
A selectable and excisable marker system for the rapid creation of recombinant poxviruses.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Oncolytic vaccinia virus immunotherapy antagonizes image-guided radiotherapy in mouse mammary tumor models.PloS one · 2024Article
- Review
- Poxvirus Recombination.Pathogens (Basel, Switzerland) · 2022Review
- The mismatched nucleotides encoded in vaccinia virus flip-and-flop hairpin telomeres serve an essential role in virion maturation.PLoS pathogens · 2022Article
- Vaccinia Virus Gene Acquisition through Nonhomologous Recombination.Journal of virology · 2021Article
- Engineering Oncolytic Vaccinia Virus to redirect Macrophages to Tumor Cells.Advances in cell and gene therapy · 2021Article
- Viral Delivery of CAR Targets to Solid Tumors Enables Effective Cell Therapy.Molecular therapy oncolytics · 2020Article
- EPPIC (Efficient Purification by Parental Inducer Constraint) Platform for Rapid Generation of Recombinant Vaccinia Viruses.Molecular therapy. Methods & clinical development · 2020Article
- Engineering and Characterization of Oncolytic Vaccinia Virus Expressing Truncated Herpes Simplex Virus Thymidine Kinase.Cancers · 2020Article
- Genes A27L and F13L as Genetic Markers for the Isolation of Recombinant Vaccinia Virus.Scientific reports · 2019Article
- Deletion of Apoptosis Inhibitor F1L in Vaccinia Virus Increases Safety and Oncolysis for Cancer Therapy.Molecular therapy oncolytics · 2019Article
- Type I IFN blockade uncouples immunotherapy-induced antitumor immunity and autoimmune toxicity.The Journal of clinical investigation · 2019Article
- Poxviruses as Gene Therapy Vectors: Generating Poxviral Vectors Expressing Therapeutic Transgenes.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Modifying the cancer-immune set point using vaccinia virus expressing re-designed interleukin-2.Nature communications · 2018Article
- Superagonist IL-15-Armed Oncolytic Virus Elicits Potent Antitumor Immunity and Therapy That Are Enhanced with PD-1 Blockade.Molecular therapy : the journal of the American Society of Gene Therapy · 2018Article
- Expression of the Vaccinia Virus Antiapoptotic F1 Protein Is Blocked by Protein Kinase R in the Absence of the Viral E3 Protein.Journal of virology · 2018Article
- Construction of an infectious horsepox virus vaccine from chemically synthesized DNA fragments.PloS one · 2018Article
- Rapid Generation of Multiple Loci-Engineered Marker-free Poxvirus and Characterization of a Clinical-Grade Oncolytic Vaccinia Virus.Molecular therapy. Methods & clinical development · 2017Article
- The vaccinia virus K7 protein promotes histone methylation associated with heterochromatin formation.PloS one · 2017Article
- Evolution of and evolutionary relationships between extant vaccinia virus strains.Journal of virology · 2015Article
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Authors and funding
9 authors.
Funding
Abstract
backgroundGenetic manipulation of poxvirus genomes through attenuation, or insertion of therapeutic genes has led to a number of vector candidates for the treatment of a variety of human diseases. The development of recombinant poxviruses often involves the genomic insertion of a selectable marker for purification and selection purposes. The use of marker genes however inevitably results in a vector that contains unwanted genetic information of no therapeutic value. METHODOLOGY/PRINCIPAL
findingsHere we describe an improved strategy that allows for the creation of marker-free recombinant poxviruses of any species. The Selectable and Excisable Marker (SEM) system incorporates a unique fusion marker gene for the efficient selection of poxvirus recombinants and the Cre/loxP system to facilitate the subsequent removal of the marker. We have defined and characterized this new methodological tool by insertion of a foreign gene into vaccinia virus, with the subsequent removal of the selectable marker. We then analyzed the importance of loxP orientation during Cre recombination, and show that the SEM system can be used to introduce site-specific deletions or inversions into the viral genome. Finally, we demonstrate that the SEM strategy is amenable to other poxviruses, as demonstrated here with the creation of an ectromelia virus recombinant lacking the EVM002 gene. CONCLUSION/SIGNIFICANCE: The system described here thus provides a faster, simpler and more efficient means to create clinic-ready recombinant poxviruses for therapeutic gene therapy applications.
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