ArticleMolecular therapy. Methods & clinical development2015
A marker-free system for highly efficient construction of vaccinia virus vectors using CRISPR Cas9.
Article in Molecular therapy. Methods & clinical development, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
31 citing papers in PubMed, 56 citations in OpenAlex.
- Cas9-Mediated Poxvirus Recombinant Recovery (CASPRR) for Fast Recovery of Recombinant Vaccinia Viruses.Methods in molecular biology (Clifton, N.J.) · 2025Article
- GM-CSF and IL-21-armed oncolytic vaccinia virus significantly enhances anti-tumor activity and synergizes with anti-PD1 immunotherapy in pancreatic cancer.Frontiers in immunology · 2024Article
- A novel oncolytic Vaccinia virus armed with IL-12 augments antitumor immune responses leading to durable regression in murine models of lung cancer.Frontiers in immunology · 2024Article
- Review
- Post-genomic era in agriculture and veterinary science: successful and proposed application of genetic targeting technologies.Frontiers in veterinary science · 2023Review
- High-throughput engineering of cytoplasmic- and nuclear-replicating large dsDNA viruses by CRISPR/Cas9.The Journal of general virology · 2022Article
- Efficient Method for Generating Point Mutations in the Vaccinia Virus Genome Using CRISPR/Cas9.Viruses · 2022Article
- Principles and Applications of CRISPR Toolkit in Virus Manipulation, Diagnosis, and Virus-Host Interactions.Cells · 2022Review
- Efficient Selection of Recombinant Fluorescent Vaccinia Virus Strains and Rapid Virus Titer Determination by Using a Multi-Well Plate Imaging System.Biomedicines · 2021Article
- MAVERICC: Marker-free Vaccinia Virus Engineering of Recombinants through in vitro CRISPR/Cas9 Cleavage.Journal of molecular biology · 2021Article
- Review
- A novel vaccinia virus enhances anti-tumor efficacy and promotes a long-term anti-tumor response in a murine model of colorectal cancer.Molecular therapy oncolytics · 2021Article
- The Application of CRISPR/Cas9 Technology for Cancer Immunotherapy: Current Status and Problems.Frontiers in oncology · 2021Review
- A systemically deliverable Vaccinia virus with increased capacity for intertumoral and intratumoral spread effectively treats pancreatic cancer.Journal for immunotherapy of cancer · 2021Article
- Rapid poxvirus engineering using CRISPR/Cas9 as a selection tool.Communications biology · 2020Article
- EPPIC (Efficient Purification by Parental Inducer Constraint) Platform for Rapid Generation of Recombinant Vaccinia Viruses.Molecular therapy. Methods & clinical development · 2020Article
- A new oncolyticJournal for immunotherapy of cancer · 2020Article
- Bioethical issues in genome editing by CRISPR-Cas9 technology.Turkish journal of biology = Turk biyoloji dergisi · 2020Article
- Application of CRISPR/Cas9 in Understanding Avian Viruses and Developing Poultry Vaccines.Frontiers in cellular and infection microbiology · 2020Review
- CRISPR-Cas based targeting of host and viral genes as an antiviral strategy.Seminars in cell & developmental biology · 2019Review
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 2 countries.
Funding
Abstract
The current method for creation of vaccinia virus (VACV) vectors involves using a selection and purification marker, however inclusion of a gene without therapeutic value in the resulting vector is not desirable for clinical use. The Cre-LoxP system has been used to make marker-free Poxviruses, but the efficiency was very low. To obtain a marker-free VACV vector, we developed marker gene excision systems to modify the thymidine kinase (TK) region and N1L regions using Cre-Loxp and Flp-FRET systems respectively. CRISPR-Cas9 system significantly resulted in a high efficiency (~90%) in generation of marker gene-positive TK-mutant VACV vector. The marker gene (RFP) could be excised from the recombinant virus using Cre recombinase. To make a marker-free VV vector with double gene deletions targeting the TK and N1L gene, we constructed a donor repair vector targeting the N1L gene, which can carry a therapeutic gene and the marker (RFP) that could be excised from the recombinant virus using Flp recombinase. The marker-free system developed here can be used to efficiently construct VACV vectors armed with any therapeutic genes in the TK region or N1L region without marker genes. Our marker-free system platform has significant potential for development of new marker-free VACV vectors for clinical application.
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.