Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development
ABSTRACT Poxviruses have large DNA genomes, and they are able to infect multiple vertebrate and invertebrate animals, including humans. Despite the eradication of smallpox, poxvirus infections still remain a significant public health concern. Vaccinia virus (VV) is the prototypic member in the poxvi...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2022-06-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.00272-22 |
| _version_ | 1811333211855257600 |
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| author | María M. Lorenzo Aitor Nogales Kevin Chiem Rafael Blasco Luis Martínez-Sobrido |
| author_facet | María M. Lorenzo Aitor Nogales Kevin Chiem Rafael Blasco Luis Martínez-Sobrido |
| author_sort | María M. Lorenzo |
| collection | DOAJ |
| description | ABSTRACT Poxviruses have large DNA genomes, and they are able to infect multiple vertebrate and invertebrate animals, including humans. Despite the eradication of smallpox, poxvirus infections still remain a significant public health concern. Vaccinia virus (VV) is the prototypic member in the poxviridae family and it has been used extensively for different prophylactic applications, including the generation of vaccines against multiple infectious diseases and/or for oncolytic treatment. Many attempts have been pursued to develop novel attenuated forms of VV with improved safety profiles for their implementation as vaccines and/or vaccines vectors. We and others have previously demonstrated how RNA viruses encoding codon-deoptimized viral genes are attenuated, immunogenic and able to protect, upon a single administration, against challenge with parental viruses. In this study, we employed the same experimental approach based on the use of misrepresented codons for the generation of a recombinant (r)VV encoding a codon-deoptimized A24R gene, which is a key component of the viral RNA polymerase. Similar to our previous studies with RNA viruses, the A24R codon-deoptimized rVV (v-A24cd) was highly attenuated in vivo but able to protect, after a single intranasal dose administration, against an otherwise lethal challenge with parental VV. These results indicate that poxviruses can be effectively attenuated by synonymous codon deoptimization and open the possibility of using this methodology alone or in combination with other experimental approaches for the development of attenuated vaccines for the treatment of poxvirus infection, or to generate improved VV-based vectors. Moreover, this approach could be applied to other DNA viruses. IMPORTANCE The family poxviridae includes multiple viruses of medical and veterinary relevance, being vaccinia virus (VV) the prototypic member in the family. VV was used during the smallpox vaccination campaign to eradicate variola virus (VARV), which is considered a credible bioterrorism threat. Because of novel innovations in genetic engineering and vaccine technology, VV has gained popularity as a viral vector for the development of vaccines against several infectious diseases. Several approaches have been used to generate attenuated VV for its implementation as vaccine and/or vaccine vector. Here, we generated a rVV containing a codon-deoptimized A24R gene (v-A24cd), which encodes a key component of the viral RNA polymerase. v-A24cd was stable in culture cells and highly attenuated in vivo but able to protect against a subsequent lethal challenge with parental VV. Our findings support the use of this approach for the development of safe, stable, and protective live-attenuated VV and/or vaccine vectors. |
| first_indexed | 2024-04-13T16:48:43Z |
| format | Article |
| id | doaj.art-be02071d740941d79a11aef164369e25 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-13T16:48:43Z |
| publishDate | 2022-06-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-be02071d740941d79a11aef164369e252022-12-22T02:39:00ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972022-06-0110310.1128/spectrum.00272-22Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine DevelopmentMaría M. Lorenzo0Aitor Nogales1Kevin Chiem2Rafael Blasco3Luis Martínez-Sobrido4Departamento de Biotecnología, Centro Nacional INIA, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, SpainDepartment of Microbiology and Immunology, University of Rochester, Rochester, New York, USADepartment of Microbiology and Immunology, University of Rochester, Rochester, New York, USADepartamento de Biotecnología, Centro Nacional INIA, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, SpainDepartment of Microbiology and Immunology, University of Rochester, Rochester, New York, USAABSTRACT Poxviruses have large DNA genomes, and they are able to infect multiple vertebrate and invertebrate animals, including humans. Despite the eradication of smallpox, poxvirus infections still remain a significant public health concern. Vaccinia virus (VV) is the prototypic member in the poxviridae family and it has been used extensively for different prophylactic applications, including the generation of vaccines against multiple infectious diseases and/or for oncolytic treatment. Many attempts have been pursued to develop novel attenuated forms of VV with improved safety profiles for their implementation as vaccines and/or vaccines vectors. We and others have previously demonstrated how RNA viruses encoding codon-deoptimized viral genes are attenuated, immunogenic and able to protect, upon a single administration, against challenge with parental viruses. In this study, we employed the same experimental approach based on the use of misrepresented codons for the generation of a recombinant (r)VV encoding a codon-deoptimized A24R gene, which is a key component of the viral RNA polymerase. Similar to our previous studies with RNA viruses, the A24R codon-deoptimized rVV (v-A24cd) was highly attenuated in vivo but able to protect, after a single intranasal dose administration, against an otherwise lethal challenge with parental VV. These results indicate that poxviruses can be effectively attenuated by synonymous codon deoptimization and open the possibility of using this methodology alone or in combination with other experimental approaches for the development of attenuated vaccines for the treatment of poxvirus infection, or to generate improved VV-based vectors. Moreover, this approach could be applied to other DNA viruses. IMPORTANCE The family poxviridae includes multiple viruses of medical and veterinary relevance, being vaccinia virus (VV) the prototypic member in the family. VV was used during the smallpox vaccination campaign to eradicate variola virus (VARV), which is considered a credible bioterrorism threat. Because of novel innovations in genetic engineering and vaccine technology, VV has gained popularity as a viral vector for the development of vaccines against several infectious diseases. Several approaches have been used to generate attenuated VV for its implementation as vaccine and/or vaccine vector. Here, we generated a rVV containing a codon-deoptimized A24R gene (v-A24cd), which encodes a key component of the viral RNA polymerase. v-A24cd was stable in culture cells and highly attenuated in vivo but able to protect against a subsequent lethal challenge with parental VV. Our findings support the use of this approach for the development of safe, stable, and protective live-attenuated VV and/or vaccine vectors.https://journals.asm.org/doi/10.1128/spectrum.00272-22vaccinia viruscodon biascodon deoptimizationA24R genelive-attenuated vaccinesGFP |
| spellingShingle | María M. Lorenzo Aitor Nogales Kevin Chiem Rafael Blasco Luis Martínez-Sobrido Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development Microbiology Spectrum vaccinia virus codon bias codon deoptimization A24R gene live-attenuated vaccines GFP |
| title | Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development |
| title_full | Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development |
| title_fullStr | Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development |
| title_full_unstemmed | Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development |
| title_short | Vaccinia Virus Attenuation by Codon Deoptimization of the A24R Gene for Vaccine Development |
| title_sort | vaccinia virus attenuation by codon deoptimization of the a24r gene for vaccine development |
| topic | vaccinia virus codon bias codon deoptimization A24R gene live-attenuated vaccines GFP |
| url | https://journals.asm.org/doi/10.1128/spectrum.00272-22 |
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