Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens

Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of genera...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Asha A. Philip [verfasserIn] Kaitlin K. Doucette [verfasserIn] Tanmaya A. Rasal [verfasserIn] John T. Patton [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	rotavirus reverse genetics vaccines enteric virus expression vector

Übergeordnetes Werk:	In: Proceedings - MDPI AG, 2018, 50(2020), 1, p 53
Übergeordnetes Werk:	volume:50 ; year:2020 ; number:1, p 53

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3390/proceedings2020050053

Katalog-ID:	DOAJ055702430

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520			\|a Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also.
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allfields	10.3390/proceedings2020050053 doi (DE-627)DOAJ055702430 (DE-599)DOAJ766585112d0e47f68ce1b37c31f2a9de DE-627 ger DE-627 rakwb eng Asha A. Philip verfasserin aut Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also. rotavirus reverse genetics vaccines enteric virus expression vector General Works A Kaitlin K. Doucette verfasserin aut Tanmaya A. Rasal verfasserin aut John T. Patton verfasserin aut In Proceedings MDPI AG, 2018 50(2020), 1, p 53 (DE-627)896671828 (DE-600)2904077-2 25043900 nnns volume:50 year:2020 number:1, p 53 https://doi.org/10.3390/proceedings2020050053 kostenfrei https://doaj.org/article/766585112d0e47f68ce1b37c31f2a9de kostenfrei https://www.mdpi.com/2504-3900/50/1/53 kostenfrei https://doaj.org/toc/2504-3900 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 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_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 50 2020 1, p 53
spelling	10.3390/proceedings2020050053 doi (DE-627)DOAJ055702430 (DE-599)DOAJ766585112d0e47f68ce1b37c31f2a9de DE-627 ger DE-627 rakwb eng Asha A. Philip verfasserin aut Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also. rotavirus reverse genetics vaccines enteric virus expression vector General Works A Kaitlin K. Doucette verfasserin aut Tanmaya A. Rasal verfasserin aut John T. Patton verfasserin aut In Proceedings MDPI AG, 2018 50(2020), 1, p 53 (DE-627)896671828 (DE-600)2904077-2 25043900 nnns volume:50 year:2020 number:1, p 53 https://doi.org/10.3390/proceedings2020050053 kostenfrei https://doaj.org/article/766585112d0e47f68ce1b37c31f2a9de kostenfrei https://www.mdpi.com/2504-3900/50/1/53 kostenfrei https://doaj.org/toc/2504-3900 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 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_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 50 2020 1, p 53
allfields_unstemmed	10.3390/proceedings2020050053 doi (DE-627)DOAJ055702430 (DE-599)DOAJ766585112d0e47f68ce1b37c31f2a9de DE-627 ger DE-627 rakwb eng Asha A. Philip verfasserin aut Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also. rotavirus reverse genetics vaccines enteric virus expression vector General Works A Kaitlin K. Doucette verfasserin aut Tanmaya A. Rasal verfasserin aut John T. Patton verfasserin aut In Proceedings MDPI AG, 2018 50(2020), 1, p 53 (DE-627)896671828 (DE-600)2904077-2 25043900 nnns volume:50 year:2020 number:1, p 53 https://doi.org/10.3390/proceedings2020050053 kostenfrei https://doaj.org/article/766585112d0e47f68ce1b37c31f2a9de kostenfrei https://www.mdpi.com/2504-3900/50/1/53 kostenfrei https://doaj.org/toc/2504-3900 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 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_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 50 2020 1, p 53
allfieldsGer	10.3390/proceedings2020050053 doi (DE-627)DOAJ055702430 (DE-599)DOAJ766585112d0e47f68ce1b37c31f2a9de DE-627 ger DE-627 rakwb eng Asha A. Philip verfasserin aut Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also. rotavirus reverse genetics vaccines enteric virus expression vector General Works A Kaitlin K. Doucette verfasserin aut Tanmaya A. Rasal verfasserin aut John T. Patton verfasserin aut In Proceedings MDPI AG, 2018 50(2020), 1, p 53 (DE-627)896671828 (DE-600)2904077-2 25043900 nnns volume:50 year:2020 number:1, p 53 https://doi.org/10.3390/proceedings2020050053 kostenfrei https://doaj.org/article/766585112d0e47f68ce1b37c31f2a9de kostenfrei https://www.mdpi.com/2504-3900/50/1/53 kostenfrei https://doaj.org/toc/2504-3900 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 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_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 50 2020 1, p 53
allfieldsSound	10.3390/proceedings2020050053 doi (DE-627)DOAJ055702430 (DE-599)DOAJ766585112d0e47f68ce1b37c31f2a9de DE-627 ger DE-627 rakwb eng Asha A. Philip verfasserin aut Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also. rotavirus reverse genetics vaccines enteric virus expression vector General Works A Kaitlin K. Doucette verfasserin aut Tanmaya A. Rasal verfasserin aut John T. Patton verfasserin aut In Proceedings MDPI AG, 2018 50(2020), 1, p 53 (DE-627)896671828 (DE-600)2904077-2 25043900 nnns volume:50 year:2020 number:1, p 53 https://doi.org/10.3390/proceedings2020050053 kostenfrei https://doaj.org/article/766585112d0e47f68ce1b37c31f2a9de kostenfrei https://www.mdpi.com/2504-3900/50/1/53 kostenfrei https://doaj.org/toc/2504-3900 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 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_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 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_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 50 2020 1, p 53
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author	Asha A. Philip
spellingShingle	Asha A. Philip misc rotavirus misc reverse genetics misc vaccines misc enteric virus misc expression vector misc General Works misc A Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens
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topic_title	Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens rotavirus reverse genetics vaccines enteric virus expression vector
topic	misc rotavirus misc reverse genetics misc vaccines misc enteric virus misc expression vector misc General Works misc A
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title	Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens
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title_full	Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens
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author_browse	Asha A. Philip Kaitlin K. Doucette Tanmaya A. Rasal John T. Patton
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title_sort	rotaviruses as neonatal vaccine expression vectors against other enteric pathogens
title_auth	Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens
abstract	Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also.
abstractGer	Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also.
abstract_unstemmed	Although the incidence of rotavirus diarrheal disease has been reduced by the introduction of neonatal rotavirus vaccines, other enteric viruses—including norovirus, hepatitis E virus (HEV), and astrovirus—remain significant causes of illness. In this study, we investigated the possibility of generating recombinant rotaviruses that express the capsid proteins of other enteric viruses as an approach for creating neonatal multitarget vaccines. As a first step, we examined whether the segmented dsRNA genome of rotavirus could be engineered to express a separate foreign protein through the use of a 2A translational “self-cleavage” element. These attempts were successful, allowing for the recovery of recombinant rotaviruses with modified-segment-7 RNAs that contained a single open reading frame (ORF) encoding a NSP3-2A-fluorescent protein (FP) cassette. By varying the FP introduced into the cassette, genetically-stable rotaviruses were generated which grew efficiently and directed the robust expression of FP as an independent product (e.g., UnaG (green), mRuby (red), mKate (orange), TagBFP (blue), and (YFP) yellow). Subsequently, attempts were made to recover recombinant rotaviruses with modified-segment-7 RNAs that contained a single ORF encoding NSP3-2A fused to the capsid-protein gene of norovirus (VP1, P, or P2), HEV (ORF2), or astrovirus (VP70 or VP90). These attempts resulted in the generation of recombinant viruses that efficiently expressed capsid proteins of other enteric viruses, despite the required addition of up to 2.5 kB of foreign sequence to the 18.5 kB rotavirus genome. Our findings support the idea that rotaviruses can be engineered as plug-and-play expression vectors to create next-generation neonatal vaccines that can induce immunological protection against not only rotavirus, but other enteric pathogens also.
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title_short	Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens
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Rotaviruses as Neonatal Vaccine Expression Vectors against Other Enteric Pathogens

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