A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes

Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared diffe...
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Gespeichert in:

Autor*in:	Xiujuan Zhao [verfasserIn] Lin Wang [verfasserIn] Qinghua Cui [verfasserIn] Ping Li [verfasserIn] Yanyan Wang [verfasserIn] Yingying Zhang [verfasserIn] Yong Yang [verfasserIn] Lijun Rong [verfasserIn] Ruikun Du [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2018

Schlagwörter:	influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication

Übergeordnetes Werk:	In: Viruses - MDPI AG, 2009, 10(2018), 12, p 679
Übergeordnetes Werk:	volume:10 ; year:2018 ; number:12, p 679

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3390/v10120679

Katalog-ID:	DOAJ044882343

Internformat


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520			\|a Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments.
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allfields	10.3390/v10120679 doi (DE-627)DOAJ044882343 (DE-599)DOAJ2f06d000a5b5419587ca3e104381dca2 DE-627 ger DE-627 rakwb eng QR1-502 Xiujuan Zhao verfasserin aut A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments. influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication Microbiology Lin Wang verfasserin aut Qinghua Cui verfasserin aut Ping Li verfasserin aut Yanyan Wang verfasserin aut Yingying Zhang verfasserin aut Yong Yang verfasserin aut Lijun Rong verfasserin aut Ruikun Du verfasserin aut In Viruses MDPI AG, 2009 10(2018), 12, p 679 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:10 year:2018 number:12, p 679 https://doi.org/10.3390/v10120679 kostenfrei https://doaj.org/article/2f06d000a5b5419587ca3e104381dca2 kostenfrei https://www.mdpi.com/1999-4915/10/12/679 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2018 12, p 679
spelling	10.3390/v10120679 doi (DE-627)DOAJ044882343 (DE-599)DOAJ2f06d000a5b5419587ca3e104381dca2 DE-627 ger DE-627 rakwb eng QR1-502 Xiujuan Zhao verfasserin aut A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments. influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication Microbiology Lin Wang verfasserin aut Qinghua Cui verfasserin aut Ping Li verfasserin aut Yanyan Wang verfasserin aut Yingying Zhang verfasserin aut Yong Yang verfasserin aut Lijun Rong verfasserin aut Ruikun Du verfasserin aut In Viruses MDPI AG, 2009 10(2018), 12, p 679 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:10 year:2018 number:12, p 679 https://doi.org/10.3390/v10120679 kostenfrei https://doaj.org/article/2f06d000a5b5419587ca3e104381dca2 kostenfrei https://www.mdpi.com/1999-4915/10/12/679 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2018 12, p 679
allfields_unstemmed	10.3390/v10120679 doi (DE-627)DOAJ044882343 (DE-599)DOAJ2f06d000a5b5419587ca3e104381dca2 DE-627 ger DE-627 rakwb eng QR1-502 Xiujuan Zhao verfasserin aut A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments. influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication Microbiology Lin Wang verfasserin aut Qinghua Cui verfasserin aut Ping Li verfasserin aut Yanyan Wang verfasserin aut Yingying Zhang verfasserin aut Yong Yang verfasserin aut Lijun Rong verfasserin aut Ruikun Du verfasserin aut In Viruses MDPI AG, 2009 10(2018), 12, p 679 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:10 year:2018 number:12, p 679 https://doi.org/10.3390/v10120679 kostenfrei https://doaj.org/article/2f06d000a5b5419587ca3e104381dca2 kostenfrei https://www.mdpi.com/1999-4915/10/12/679 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2018 12, p 679
allfieldsGer	10.3390/v10120679 doi (DE-627)DOAJ044882343 (DE-599)DOAJ2f06d000a5b5419587ca3e104381dca2 DE-627 ger DE-627 rakwb eng QR1-502 Xiujuan Zhao verfasserin aut A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments. influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication Microbiology Lin Wang verfasserin aut Qinghua Cui verfasserin aut Ping Li verfasserin aut Yanyan Wang verfasserin aut Yingying Zhang verfasserin aut Yong Yang verfasserin aut Lijun Rong verfasserin aut Ruikun Du verfasserin aut In Viruses MDPI AG, 2009 10(2018), 12, p 679 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:10 year:2018 number:12, p 679 https://doi.org/10.3390/v10120679 kostenfrei https://doaj.org/article/2f06d000a5b5419587ca3e104381dca2 kostenfrei https://www.mdpi.com/1999-4915/10/12/679 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2018 12, p 679
allfieldsSound	10.3390/v10120679 doi (DE-627)DOAJ044882343 (DE-599)DOAJ2f06d000a5b5419587ca3e104381dca2 DE-627 ger DE-627 rakwb eng QR1-502 Xiujuan Zhao verfasserin aut A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments. influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication Microbiology Lin Wang verfasserin aut Qinghua Cui verfasserin aut Ping Li verfasserin aut Yanyan Wang verfasserin aut Yingying Zhang verfasserin aut Yong Yang verfasserin aut Lijun Rong verfasserin aut Ruikun Du verfasserin aut In Viruses MDPI AG, 2009 10(2018), 12, p 679 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:10 year:2018 number:12, p 679 https://doi.org/10.3390/v10120679 kostenfrei https://doaj.org/article/2f06d000a5b5419587ca3e104381dca2 kostenfrei https://www.mdpi.com/1999-4915/10/12/679 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2018 12, p 679
language	English
source	In Viruses 10(2018), 12, p 679 volume:10 year:2018 number:12, p 679
sourceStr	In Viruses 10(2018), 12, p 679 volume:10 year:2018 number:12, p 679
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication Microbiology
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container_title	Viruses
authorswithroles_txt_mv	Xiujuan Zhao @@aut@@ Lin Wang @@aut@@ Qinghua Cui @@aut@@ Ping Li @@aut@@ Yanyan Wang @@aut@@ Yingying Zhang @@aut@@ Yong Yang @@aut@@ Lijun Rong @@aut@@ Ruikun Du @@aut@@
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callnumber-first	Q - Science
author	Xiujuan Zhao
spellingShingle	Xiujuan Zhao misc QR1-502 misc influenza A virus misc reporter virus misc attenuation misc mechanism misc noninfectious particles misc vRNA replication misc Microbiology A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes
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topic_title	QR1-502 A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes influenza A virus reporter virus attenuation mechanism noninfectious particles vRNA replication
topic	misc QR1-502 misc influenza A virus misc reporter virus misc attenuation misc mechanism misc noninfectious particles misc vRNA replication misc Microbiology
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title	A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes
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title_sort	mechanism underlying attenuation of recombinant influenza a viruses carrying reporter genes
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title_auth	A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes
abstract	Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments.
abstractGer	Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments.
abstract_unstemmed	Influenza A viruses (IAV) carrying reporter genes provide a powerful tool to study viral infection and pathogenesis in vivo, however, incorporating a non-essential gene into the IAV genome often results in virus attenuation and genetic instability. Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. In summary, we unravel a mechanism underlying attenuation of reporter IAVs, which suggests a new approach to restore infectivity and virulence by introducing extra mutations compensating for the impaired replication property of corresponding segments.
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title_short	A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes
url	https://doi.org/10.3390/v10120679 https://doaj.org/article/2f06d000a5b5419587ca3e104381dca2 https://www.mdpi.com/1999-4915/10/12/679 https://doaj.org/toc/1999-4915
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up_date	2024-07-04T00:50:45.846Z
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Very few studies have systematically compared different reporter IAVs, and most optimization attempts seem to lack authentic directions. In this study, we evaluated the ratio of genome copies to the number of infectious unit of two reporter IAVs, PR8-NS1-Gluc and PR8-PB2-Gluc. As a result, PR8-NS1-Gluc and PR8-PB2-Gluc produced 41.4 and 3.8 genomes containing noninfectious particles respectively for every such particle produced by parental PR8 virus. RdRp assay demonstrated that modification of segment NS by inserting reporter genes can interfere with the replication competitive property of the corresponding vRNAs, and the balance of the 8 segments of the reporter IAVs were drastically impaired in infected cells. As a consequence, large amounts of NS-null noninfectious particles were produced during the PR8-NS1-Gluc packaging. 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A Mechanism Underlying Attenuation of Recombinant Influenza A Viruses Carrying Reporter Genes

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