Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most challenging infectious disease of pig populations causing devastating economic loss to swine industry. Reverse genetics allow to engineer modified viruses such attenuated strains for vaccine development. Some reverse gene...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2021-01-01
|
Series: | Computational and Structural Biotechnology Journal |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037021003718 |
id |
doaj-31e076c516df43dd92c71d9fc7d1dd45 |
---|---|
record_format |
Article |
spelling |
doaj-31e076c516df43dd92c71d9fc7d1dd452021-09-21T04:09:03ZengElsevierComputational and Structural Biotechnology Journal2001-03702021-01-011951085116Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragmentsJulien Mélade0Géraldine Piorkowski1Hawa Sophia Bouzidi2Alain Medawar3Claudine Raffy4Xavier de Lamballerie5Antoine Nougairède6Unité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207), Marseille, France; Corresponding author at: Faculté de Médecine de Marseille 27, Bd Jean Moulin, 13005 Marseille, France.Unité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207), Marseille, FranceUnité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207), Marseille, France; VIRBAC, 1e Avenue, 13ème rue, LID, BP27 - 06511 Carros, FranceVIRBAC, 1e Avenue, 13ème rue, LID, BP27 - 06511 Carros, FranceVIRBAC, 1e Avenue, 13ème rue, LID, BP27 - 06511 Carros, FranceUnité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207), Marseille, FranceUnité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207), Marseille, FrancePorcine reproductive and respiratory syndrome virus (PRRSV) is one of the most challenging infectious disease of pig populations causing devastating economic loss to swine industry. Reverse genetics allow to engineer modified viruses such attenuated strains for vaccine development. Some reverse genetic systems were described for PRRSVs but, due to genome complexity of PRRSVs, construction and modification of such systems remain laborious and time-consuming.In this study, we described a reverse genetics approach based on the “Infectious-Subgenomic Amplicons” (ISA) method to rescue infectious PRRSV particles. Permissive cells were transfected with 4 overlapping synthetic DNA fragments covering the entire genome of PRRSV which allowed the rapid reconstruction of the complete virus genome and the subsequent generation of infectious wild-type particles within days.The ISA method represent a rapid alternative of conventional reverse genetic systems. This method will help to generate genetically modified and attenuated strains for the development of sanitary countermeasures in the future.http://www.sciencedirect.com/science/article/pii/S2001037021003718PRRSVReverse geneticssynthetic DNA fragmentsISA |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Julien Mélade Géraldine Piorkowski Hawa Sophia Bouzidi Alain Medawar Claudine Raffy Xavier de Lamballerie Antoine Nougairède |
spellingShingle |
Julien Mélade Géraldine Piorkowski Hawa Sophia Bouzidi Alain Medawar Claudine Raffy Xavier de Lamballerie Antoine Nougairède Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments Computational and Structural Biotechnology Journal PRRSV Reverse genetics synthetic DNA fragments ISA |
author_facet |
Julien Mélade Géraldine Piorkowski Hawa Sophia Bouzidi Alain Medawar Claudine Raffy Xavier de Lamballerie Antoine Nougairède |
author_sort |
Julien Mélade |
title |
Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments |
title_short |
Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments |
title_full |
Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments |
title_fullStr |
Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments |
title_full_unstemmed |
Rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic DNA fragments |
title_sort |
rapid reconstruction of porcine reproductive and respiratory syndrome virus using synthetic dna fragments |
publisher |
Elsevier |
series |
Computational and Structural Biotechnology Journal |
issn |
2001-0370 |
publishDate |
2021-01-01 |
description |
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most challenging infectious disease of pig populations causing devastating economic loss to swine industry. Reverse genetics allow to engineer modified viruses such attenuated strains for vaccine development. Some reverse genetic systems were described for PRRSVs but, due to genome complexity of PRRSVs, construction and modification of such systems remain laborious and time-consuming.In this study, we described a reverse genetics approach based on the “Infectious-Subgenomic Amplicons” (ISA) method to rescue infectious PRRSV particles. Permissive cells were transfected with 4 overlapping synthetic DNA fragments covering the entire genome of PRRSV which allowed the rapid reconstruction of the complete virus genome and the subsequent generation of infectious wild-type particles within days.The ISA method represent a rapid alternative of conventional reverse genetic systems. This method will help to generate genetically modified and attenuated strains for the development of sanitary countermeasures in the future. |
topic |
PRRSV Reverse genetics synthetic DNA fragments ISA |
url |
http://www.sciencedirect.com/science/article/pii/S2001037021003718 |
work_keys_str_mv |
AT julienmelade rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments AT geraldinepiorkowski rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments AT hawasophiabouzidi rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments AT alainmedawar rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments AT claudineraffy rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments AT xavierdelamballerie rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments AT antoinenougairede rapidreconstructionofporcinereproductiveandrespiratorysyndromevirususingsyntheticdnafragments |
_version_ |
1717373787278671872 |