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Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein

Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the Arteriviridae family, is one of the most common and economically important swine pathogens. Although both live-attenuated and killed-inactivated vaccines against the virus have been available for a decade, PRRSV is still a...

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Autores principales: Song, Byung-Hak, Kim, Jeong-Min, Kim, Jin-Kyoung, Jang, Han-Saem, Yun, Gil-Nam, Choi, Eun-Jin, Song, Jae-Young, Yun, Sang-Im, Lee, Young-Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Microbiological Society of Korea 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7091078/
https://www.ncbi.nlm.nih.gov/pubmed/21717343
http://dx.doi.org/10.1007/s12275-011-1280-1
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author Song, Byung-Hak
Kim, Jeong-Min
Kim, Jin-Kyoung
Jang, Han-Saem
Yun, Gil-Nam
Choi, Eun-Jin
Song, Jae-Young
Yun, Sang-Im
Lee, Young-Min
author_facet Song, Byung-Hak
Kim, Jeong-Min
Kim, Jin-Kyoung
Jang, Han-Saem
Yun, Gil-Nam
Choi, Eun-Jin
Song, Jae-Young
Yun, Sang-Im
Lee, Young-Min
author_sort Song, Byung-Hak
collection PubMed
description Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the Arteriviridae family, is one of the most common and economically important swine pathogens. Although both live-attenuated and killed-inactivated vaccines against the virus have been available for a decade, PRRSV is still a major problem in the swine industry worldwide. To explore the possibility of producing single-round infectious PRRSV replicon particles as a potential vaccine strategy, we have now generated two necessary components: 1) a stable cell line (BHK/Sinrepl9/PRRSV-N) that constitutively expresses the viral nucleocapsid (N) protein localized to the cytoplasm and the nucleolus and 2) a PRRSV replicon vector (pBAC/PRRSV/Replicon-AN) with a 177-nucleotide deletion, removing the 3′-half portion of ORF7 in the viral genome, from which the self-replicating propagation-defective replicon RNAs were synthesized in vitro by SP6 polymerase run-off transcription. Transfection of this replicon RNA into N protein-expressing BHK-21 cells led to the secretion of infectious particles that packaged the replicon RNA, albeit with a low production efficiency of 0.4 × 10(2) to 1.1 × 10(2) infectious units/ml; the produced particles had only single-round infectivity with no cell-to-cell spread. This trans-complementation system for PRRSV provides a useful platform for studies to define the packaging signals and motifs present within the viral genome and N protein, respectively, and to develop viral replicon-based antiviral vaccines that will stop the infection and spread of this pathogen.
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spelling pubmed-70910782020-03-24 Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein Song, Byung-Hak Kim, Jeong-Min Kim, Jin-Kyoung Jang, Han-Saem Yun, Gil-Nam Choi, Eun-Jin Song, Jae-Young Yun, Sang-Im Lee, Young-Min J Microbiol Note Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the Arteriviridae family, is one of the most common and economically important swine pathogens. Although both live-attenuated and killed-inactivated vaccines against the virus have been available for a decade, PRRSV is still a major problem in the swine industry worldwide. To explore the possibility of producing single-round infectious PRRSV replicon particles as a potential vaccine strategy, we have now generated two necessary components: 1) a stable cell line (BHK/Sinrepl9/PRRSV-N) that constitutively expresses the viral nucleocapsid (N) protein localized to the cytoplasm and the nucleolus and 2) a PRRSV replicon vector (pBAC/PRRSV/Replicon-AN) with a 177-nucleotide deletion, removing the 3′-half portion of ORF7 in the viral genome, from which the self-replicating propagation-defective replicon RNAs were synthesized in vitro by SP6 polymerase run-off transcription. Transfection of this replicon RNA into N protein-expressing BHK-21 cells led to the secretion of infectious particles that packaged the replicon RNA, albeit with a low production efficiency of 0.4 × 10(2) to 1.1 × 10(2) infectious units/ml; the produced particles had only single-round infectivity with no cell-to-cell spread. This trans-complementation system for PRRSV provides a useful platform for studies to define the packaging signals and motifs present within the viral genome and N protein, respectively, and to develop viral replicon-based antiviral vaccines that will stop the infection and spread of this pathogen. The Microbiological Society of Korea 2011-06-30 2011 /pmc/articles/PMC7091078/ /pubmed/21717343 http://dx.doi.org/10.1007/s12275-011-1280-1 Text en © The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg 2011 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Note
Song, Byung-Hak
Kim, Jeong-Min
Kim, Jin-Kyoung
Jang, Han-Saem
Yun, Gil-Nam
Choi, Eun-Jin
Song, Jae-Young
Yun, Sang-Im
Lee, Young-Min
Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein
title Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein
title_full Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein
title_fullStr Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein
title_full_unstemmed Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein
title_short Packaging of porcine reproductive and respiratory syndrome virus replicon RNA by a stable cell line expressing its nucleocapsid protein
title_sort packaging of porcine reproductive and respiratory syndrome virus replicon rna by a stable cell line expressing its nucleocapsid protein
topic Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7091078/
https://www.ncbi.nlm.nih.gov/pubmed/21717343
http://dx.doi.org/10.1007/s12275-011-1280-1
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