ArticleCommunications biology2020
Rapid poxvirus engineering using CRISPR/Cas9 as a selection tool.
Article in Communications biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Antibiotic-mediated selection of randomly mutagenized and cytokine-expressing oncolytic viruses.Nature biomedical engineering · 2025Article
- A synthetic genomics-based African swine fever virus engineering platform.Science advances · 2025Article
- Article
- Cas9-Mediated Poxvirus Recombinant Recovery (CASPRR) for Fast Recovery of Recombinant Vaccinia Viruses.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Rapid Generation of Recombinant Poxviruses Using CRISPR/Cas9 Gene Editing.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Review
- High-throughput engineering of cytoplasmic- and nuclear-replicating large dsDNA viruses by CRISPR/Cas9.The Journal of general virology · 2022Article
- Poxvirus Recombination.Pathogens (Basel, Switzerland) · 2022Review
- Efficient Method for Generating Point Mutations in the Vaccinia Virus Genome Using CRISPR/Cas9.Viruses · 2022Article
- Efficient production of protein complexes in mammalian cells using a poxvirus vector.PloS one · 2022Article
- CRISPR-mediated rapid arming of poxvirus vectors enables facile generation of the novel immunotherapeutic STINGPOX.Frontiers in immunology · 2022Article
- Selection of Vaccinia Virus Recombinants Using CRISPR/Cas9.Bio-protocol · 2021Article
- Advancements in the Growth and Construction of Recombinant Lumpy Skin Disease Virus (LSDV) for Use as a Vaccine Vector.Vaccines · 2021Article
- Article
- MAVERICC: Marker-free Vaccinia Virus Engineering of Recombinants through in vitro CRISPR/Cas9 Cleavage.Journal of molecular biology · 2021Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In standard uses of CRISPR/Cas9 technology, the cutting of genomes and their efficient repair are considered to go hand-in-hand to achieve desired genetic changes. This includes the current approach for engineering genomes of large dsDNA viruses. However, for poxviruses we show that Cas9-guide RNA complexes cut viral genomes soon after their entry into cells, but repair of these breaks is inefficient. As a result, Cas9 targeting makes only modest, if any, improvements to basal rates of homologous recombination between repair constructs and poxvirus genomes. Instead, Cas9 cleavage leads to inhibition of poxvirus DNA replication thereby suppressing virus spread in culture. This unexpected outcome allows Cas9 to be used as a powerful tool for selecting conventionally generated poxvirus recombinants, which are otherwise impossible to separate from a large background of parental virus without the use of marker genes. This application of CRISPR/Cas9 greatly speeds up the generation of poxvirus-based vaccines, making this platform considerably more attractive in the context of personalised cancer vaccines and emerging disease outbreaks.
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