A simple method for rapid cloning of complete herpesvirus genomes

Knickmann, Jan; Staliunaite, Laura; Puhach, Olha; Ostermann, Eleonore; Günther, Thomas; Nichols, Jenna; Jarvis, Michael A.; Voigt, Sebastian; Grundhoff, Adam; Davison, Andrew J.; Brune, Wolfram

doi:10.1016/j.crmeth.2024.100696

Cited by 8 publications

(1 citation statement)

References 41 publications

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“…However, the BAC and fosmid strategies must avoid the laborious steps of constructing plasmids that carry viral genomes. In recent months, a simple, rapid, and efficient method based on single-step transformation-associated recombination (STAR) in Saccharomyces cerevisiae was used to generate complete betaherpesvirus and gammaherpesvirus genomes [ 37 ]. Conveniently, the linear viral genome can be directly integrated into a vector comprising a yeast centromeric plasmid and a BAC replicon, which avoids the laborious steps of generating plasmids carrying viral genomes.…”

Section: Various Important Virus Vectorsmentioning

confidence: 99%

Current Status of Poultry Recombinant Virus Vector Vaccine Development

Wang,

Tian,

Zhao

et al. 2024

Vaccines

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Inactivated and live attenuated vaccines are the mainstays of preventing viral poultry diseases. However, the development of recombinant DNA technology in recent years has enabled the generation of recombinant virus vector vaccines, which have the advantages of preventing multiple diseases simultaneously and simplifying the vaccination schedule. More importantly, some can induce a protective immune response in the presence of maternal antibodies and offer long-term immune protection. These advantages compensate for the shortcomings of traditional vaccines. This review describes the construction and characterization of primarily poultry vaccine vectors, including fowl poxvirus (FPV), fowl adenovirus (FAdV), Newcastle disease virus (NDV), Marek’s disease virus (MDV), and herpesvirus of turkey (HVT). In addition, the pathogens targeted and the immunoprotective effect of different poultry recombinant virus vector vaccines are also presented. Finally, this review discusses the challenges in developing vector vaccines and proposes strategies for improving immune efficacy.

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Section: Various Important Virus Vectorsmentioning

confidence: 99%

Current Status of Poultry Recombinant Virus Vector Vaccine Development

Wang,

Tian,

Zhao

et al. 2024

Vaccines

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Efficient Strategy for Synthesizing Vector-Free and Oncolytic Herpes Simplex Type 1 Viruses

Xiao,

Hu,

Guo

et al. 2024

ACS Synth. Biol.

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Synthesizing viral genomes plays an important role in fundamental virology research and in the development of vaccines and antiviral drugs. Herpes simplex virus type 1 (HSV-1) is a large DNA virus widely used in oncolytic virotherapy. Although de novo synthesis of the HSV-1 genome has been previously reported, the synthetic procedure is still far from efficient, and the synthesized genome contains a vector sequence that may affect its replication and application. In the present study, we developed an efficient vector-free strategy for synthesis and rescue of synthetic HSV-1. In contrast to the conventional method of transfecting mammalian cells with a completely synthesized genome containing a vector, overlapping HSV-1 fragments synthesized by transformation-associated recombination (TAR) in yeast were linearized and cotransfected into mammalian cells to rescue the synthetic virus. Using this strategy, a synthetic virus, F-Syn, comprising the complete genome of the HSV-1 F strain, was generated. The growth curve and electron microscopy of F-Syn confirmed that its replication dynamics and morphogenesis are similar to those of the parental virus. In addition, by combining TAR with in vitro CRISPR/Cas9 editing, an oncolytic virus, F-Syn-O, with deleted viral genes ICP6, ICP34.5, and ICP47 was generated. The antitumor effect of F-Syn-O was tested in vitro. F-Syn-O established a successful infection and induced dose-dependent cytotoxic effects in various human tumor cell lines. These strategies will facilitate convenient and systemic manipulation of HSV-1 genomes and could be further applied to the design and construction of oncolytic herpesviruses.

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Human cytomegalovirus glycoprotein variants governing viral tropism and syncytium formation in epithelial cells and macrophages

Cimato,

Zhou,

Brune

et al. 2024

Preprint

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Human cytomegalovirus (HCMV) displays a broad cell tropism, and the infection of biologically relevant cells such as epithelial, endothelial, and hematopoietic cells supports viral transmission, systemic spread, and pathogenesis in the human host. HCMV strains differ in their ability to infect and replicate in these cell types, but the genetic basis of these differences has remained incompletely understood. In this study, we investigated HCMV strain VR1814, which is highly infectious for epithelial cells and macrophages and induces cell-cell fusion in both cell types. A VR1814-derived bacterial artificial chromosome (BAC) clone, FIX-BAC, was generated many years ago but has fallen out of favor because of its modest infectivity. By sequence comparison and genetic engineering of FIX, we demonstrate that the high infectivity of VR1814 and its ability to induce syncytium formation in epithelial cells and macrophages depends on VR1814-specific variants of the envelope glycoproteins gB, UL128, and UL130. We also show that UL130-neutralizing antibodies inhibit syncytium formation, and a FIX-specific mutation in UL130 is responsible for its low infectivity by reducing the amount of the pentameric glycoprotein complex in viral particles. Moreover, we found that a VR1814-specific mutation in US28 further increases viral infectivity in macrophages, possibly by promoting lytic rather than latent infection of these cells. Our findings show that variants of gB and the pentameric complex are major determinants of infectivity and syncytium formation in epithelial cells and macrophages. Furthermore, the VR1814-adjusted FIX strains can serve as valuable tools to study HCMV infection of myeloid cells.

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A simple method for rapid cloning of complete herpesvirus genomes

Cited by 8 publications

References 41 publications

Current Status of Poultry Recombinant Virus Vector Vaccine Development

Current Status of Poultry Recombinant Virus Vector Vaccine Development

Efficient Strategy for Synthesizing Vector-Free and Oncolytic Herpes Simplex Type 1 Viruses

Human cytomegalovirus glycoprotein variants governing viral tropism and syncytium formation in epithelial cells and macrophages

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