Production of therapeutic iduronate‐2‐sulfatase enzyme with a novel single‐stranded RNA virus vector

2021 ◽  
Author(s):  
Mari Ohira ◽  
Emika Kikuchi ◽  
Shiori Mizuta ◽  
Naomi Yoshida ◽  
Masafumi Onodera ◽  
...  
Keyword(s):  
Rna Virus ◽  
Virus Vector

scholarly journals Development of a Honey Bee RNA Virus Vector Based on the Genome of Deformed Wing Virus

2020 ◽  
Author(s):  
Eugene V. Ryabov ◽  
Krisztina Christmon ◽  
Matthew C. Heerman ◽  
Francisco Posada-Florez ◽  
Robert L. Harrison ◽  
...  
Keyword(s):  
Honey Bee ◽  
Fluorescent Protein ◽  
Essential Gene ◽  
Rna Virus ◽  
Natural Infection ◽  
Fluorescent Microscopy ◽  
Virus Vector ◽  
Cdna Clones ◽  
Viral Pathogen ◽  
Deformed Wing Virus

We developed a honey bee RNA-virus vector based on the genome of a picorna-like Deformed wing virus (DWV), the main viral pathogen of the honey bee (Apis mellifera). To test the potential of DWV to be utilized as a vector, the 717 nt sequence coding for the enhanced green fluorescent protein (eGFP), flanked by the peptides targeted by viral protease, was inserted into an infectious cDNA clone of DWV in-frame between the leader protein and the virus structural protein VP2 genes. The in vitro RNA transcripts from egfp-tagged DWV cDNA clones were infectious when injected into honey bee pupae. Stable DWV particles containing genomic RNA of the recovered DWV with egfp inserts were produced, as evidenced by cesium chloride density gradient centrifugation. These particles were infectious to honey bee pupae when injected intra-abdominally. Fluorescent microscopy showed GFP expression in the infected cells and Western blot analysis demonstrated accumulation of free eGFP rather than its fusions with DWV LP and/or VP2 proteins. Analysis of the progeny egfp-tagged DWV showed gradual accumulation of genome deletions for egfp, providing estimates for the rate of loss of a non-essential gene an insect RNA virus genome during natural infection.

scholarly journals Expression of the Surface Glycoproteins of Human Parainfluenza Virus Type 3 by Bovine Parainfluenza Virus Type 3, a Novel Attenuated Virus Vaccine Vector

2000 ◽  
Vol 74 (24) ◽  
pp. 11626-11635 ◽  
Author(s):  
Aurelia A. Haller ◽  
Tessa Miller ◽  
Misrach Mitiku ◽  
Kathleen Coelingh
Keyword(s):  
Virus Type ◽  
Rna Virus ◽  
Vero Cells ◽  
Virus Vector ◽  
Parainfluenza Virus ◽  
Temperature Sensitive ◽  
Young Infants ◽  
Human Parainfluenza Virus ◽  
Type 3 ◽  
Bovine Parainfluenza Virus

ABSTRACT Bovine parainfluenza virus type 3 (bPIV3) is being evaluated as an intranasal vaccine for protection against human PIV3 (hPIV3). In young infants, the bPIV3 vaccine appears to be infectious, attenuated, immunogenic, and genetically stable, which are desirable characteristics for an RNA virus vector. To test the potential of the bPIV3 vaccine strain as a vector, an infectious DNA clone of bPIV3 was assembled and recombinant bPIV3 (r-bPIV3) was rescued. r-bPIV3 displayed a temperature-sensitive phenotype for growth in tissue culture at 39°C and was attenuated in the lungs of Syrian golden hamsters. In order to test whether r-bPIV3 could serve as a vector, the fusion and hemagglutinin-neuraminidase genes of bPIV3 were replaced with those of hPIV3. The resulting bovine/human PIV3 was temperature sensitive for growth in Vero cells at 37°C. The replication of bovine/human PIV3 was also restricted in the lungs of hamsters, albeit not as severely as was observed for r-bPIV3. Despite the attenuation phenotypes observed for r-bPIV3 and bovine/human PIV3, both of these viruses protected hamsters completely upon challenge with hPIV3. In summary, bPIV3 was shown to function as a virus vector that may be especially suitable for vaccination of infants and children against PIV3 and other viruses.

scholarly journals Development of a honey bee RNA virus vector based on the genome of Deformed wing virus

2020 ◽  
Author(s):  
Eugene V. Ryabov ◽  
Krisztina Christmon ◽  
Matthew C. Heerman ◽  
Francisco Posada-Florez ◽  
Robert L. Harrison ◽  
...  
Keyword(s):  
Honey Bee ◽  
Fluorescent Protein ◽  
Essential Gene ◽  
Rna Virus ◽  
Natural Infection ◽  
Fluorescent Microscopy ◽  
Virus Vector ◽  
Cdna Clones ◽  
Viral Pathogen ◽  
Deformed Wing Virus

AbstractWe developed a honey bee RNA-virus vector based on the genome of a picorna-like Deformed wing virus (DWV), the main viral pathogen of the honey bee (Apis mellifera). To test the potential of DWV to be utilized as a vector, the 717 nt sequence coding for the enhanced green fluorescent protein (eGFP), flanked by the peptides targeted by viral protease, was inserted into an infectious cDNA clone of DWV in-frame between the leader protein and the virus structural protein VP2 genes. The in vitro RNA transcripts from egfp-tagged DWV cDNA clones were infectious when injected into honey bee pupae. Stable DWV particles containing genomic RNA of the recovered DWV with egfp inserts were produced, as evidenced by cesium chloride density gradient centrifugation. These particles were infectious to honey bee pupae when injected intra-abdominally. Fluorescent microscopy showed GFP expression in the infected cells and Western blot analysis demonstrated accumulation of free eGFP rather than its fusions with DWV LP and/or VP2 proteins. Analysis of the progeny egfp-tagged DWV showed gradual accumulation of genome deletions for egfp, providing estimates for the rate of loss of a non-essential gene an insect RNA virus genome during natural infection.

scholarly journals Viral and host determinants of RNA virus vector replication and expression

Vaccine ◽  
2005 ◽  
Vol 23 (15) ◽  
pp. 1784-1787 ◽  
Author(s):  
Paul Ahlquist ◽  
Michael Schwartz ◽  
Jianbo Chen ◽  
David Kushner ◽  
Linhui Hao ◽  
...  
Keyword(s):  
Rna Virus ◽  
Virus Vector ◽  
Host Determinants

scholarly journals Development of a Honey Bee RNA Virus Vector Based on the Genome of a Deformed Wing Virus

Viruses ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 374 ◽  
Author(s):  
Eugene V. Ryabov ◽  
Krisztina Christmon ◽  
Matthew C. Heerman ◽  
Francisco Posada-Florez ◽  
Robert L. Harrison ◽  
...  
Keyword(s):  
Honey Bee ◽  
Essential Gene ◽  
Rna Virus ◽  
Natural Infection ◽  
Fluorescent Microscopy ◽  
Virus Vector ◽  
Cdna Clones ◽  
Viral Pathogen ◽  
Deformed Wing Virus ◽  
Leader Protein

We developed a honey bee RNA-virus vector based on the genome of a picorna-like Deformed wing virus (DWV), the main viral pathogen of the honey bee (Apis mellifera). To test the potential of DWV to be utilized as a vector, the 717 nt sequence coding for the enhanced green fluorescent protein (eGFP), flanked by the peptides targeted by viral protease, was inserted into an infectious cDNA clone of DWV in-frame between the leader protein and the virus structural protein VP2 genes. The in vitro RNA transcripts from egfp-tagged DWV cDNA clones were infectious when injected into honey bee pupae. Stable DWV particles containing genomic RNA of the recovered DWV with egfp inserts were produced, as evidenced by cesium chloride density gradient centrifugation. These particles were infectious to honey bee pupae when injected intra-abdominally. Fluorescent microscopy showed GFP expression in the infected cells and Western blot analysis demonstrated accumulation of free eGFP rather than its fusions with DWV leader protein (LP) and/or viral protein (VP) 2. Analysis of the progeny egfp-tagged DWV showed gradual accumulation of genome deletions for egfp, providing estimates for the rate of loss of a non-essential gene an insect RNA virus genome during natural infection.

The Three-dimensional structure of frozen-hydrated nudaurelia capensis β virus

1987 ◽  
Vol 45 ◽  
pp. 650-651
Author(s):  
N. H. Olson ◽  
T. S. Baker ◽  
Wu Bo Mu ◽  
J. E. Johnson ◽  
D. A. Hendry
Keyword(s):  
South African ◽  
Rna Virus ◽  
Carbon Film ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Electron Dose ◽  
Total Electron ◽  
Three Dimensional Structure ◽  
Negative Stain ◽  
Dimensional Reconstruction

Nudaurelia capensis β virus (NβV) is an RNA virus of the South African Pine Emperor moth, Nudaurelia cytherea capensis (Lepidoptera: Saturniidae). The NβV capsid is a T = 4 icosahedron that contains 60T = 240 subunits of the coat protein (Mr = 61,000). A three-dimensional reconstruction of the NβV capsid was previously computed from visions embedded in negative stain suspended over holes in a carbon film. We have re-examined the three-dimensional structure of NβV, using cryo-microscopy to examine the native, unstained structure of the virion and to provide a initial phasing model for high-resolution x-ray crystallographic studiesNβV was purified and prepared for cryo-microscopy as described. Micrographs were recorded ∼1 - 2 μm underfocus at a magnification of 49,000X with a total electron dose of about 1800 e-/nm2.

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