New Advances on Zika Virus Research

Martínez-Sobrido, Luis; Almazán, Fernando

doi:10.3390/v11030258

Cited by 4 publications

(4 citation statements)

References 35 publications

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“…Assembly of the SARS-CoV-2 genome in the BAC. We used a BAC-based approach, similar to the one that we previously described for Zika virus (16)(17)(18)(19)(20)(21), to assemble an infectious clone of SARS-CoV-2 based on the USA-WA1/2020 strain ( Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome

Chiem

Park

et al. 2020

mBio

Self Cite

109

121

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Infectious coronavirus (CoV) disease 2019 (COVID-19) emerged in the city of Wuhan (China) in December 2019, causing a pandemic that has dramatically impacted public health and socioeconomic activities worldwide. A previously unknown coronavirus, severe acute respiratory syndrome CoV-2 (SARS-CoV-2), has been identified as the causative agent of COVID-19. To date, there are no U.S. Food and Drug Administration (FDA)-approved vaccines or therapeutics available for the prevention or treatment of SARS-CoV-2 infection and/or associated COVID-19 disease, which has triggered a large influx of scientific efforts to develop countermeasures to control SARS-CoV-2 spread. To contribute to these efforts, we have developed an infectious cDNA clone of the SARS-CoV-2 USA-WA1/2020 strain based on the use of a bacterial artificial chromosome (BAC). Recombinant SARS-CoV-2 (rSARS-CoV-2) was readily rescued by transfection of the BAC into Vero E6 cells. Importantly, BAC-derived rSARS-CoV-2 exhibited growth properties and plaque sizes in cultured cells comparable to those of the natural SARS-CoV-2 isolate. Likewise, rSARS-CoV-2 showed levels of replication similar to those of the natural isolate in nasal turbinates and lungs of infected golden Syrian hamsters. This is, to our knowledge, the first BAC-based reverse genetics system for the generation of infectious rSARS-CoV-2 that displays features in vivo similar to those of a natural viral isolate. This SARS-CoV-2 BAC-based reverse genetics will facilitate studies addressing several important questions in the biology of SARS-CoV-2, as well as the identification of antivirals and development of vaccines for the treatment of SARS-CoV-2 infection and associated COVID-19 disease. IMPORTANCE The pandemic coronavirus (CoV) disease 2019 (COVID-19) caused by severe acute respiratory syndrome CoV-2 (SARS-CoV-2) is a major threat to global human health. To date, there are no approved prophylactics or therapeutics available for COVID-19. Reverse genetics is a powerful approach to understand factors involved in viral pathogenesis, antiviral screening, and vaccine development. In this study, we describe the feasibility of generating recombinant SARS-CoV-2 (rSARS-CoV-2) by transfection of a single bacterial artificial chromosome (BAC). Importantly, rSARS-CoV-2 possesses the same phenotype as the natural isolate in vitro and in vivo. This is the first description of a BAC-based reverse genetics system for SARS-CoV-2 and the first time that an rSARS-CoV-2 isolate has been shown to be phenotypically identical to a natural isolate in a validated animal model of SARS-CoV-2 infection. The BAC-based reverse genetics approach will facilitate the study of SARS-CoV-2 and the development of prophylactics and therapeutics for the treatment of COVID-19.

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Section: Resultsmentioning

confidence: 99%

Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome

Chiem

Park

et al. 2020

mBio

Self Cite

109

121

View full text Add to dashboard Cite

show abstract

“…We used a BAC-based approach, similar to the one we previously described for Zika virus (ZIKV) (16)(17)(18)(19)(20)(21) to assemble an infectious clone of SARS-CoV-2 based on the USA-WA1/2020 strain (Fig. 1A).…”

Section: Assembly Of Sars-cov-2 Genome In the Bacmentioning

confidence: 99%

Rescue of SARS-CoV-2 from a single bacterial artificial chromosome

Chiem

Park

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

An infectious coronavirus disease 2019 (COVID-19) emerged in the city of Wuhan (China) in December 2019, causing a pandemic that has dramatically impacted public health and socioeconomic activities worldwide. A previously unknown coronavirus, Severe Acute Respiratory Syndrome CoV-2 (SARS-CoV-2), has been identified as the causative agent of COVID-19. To date, there are no United States (US) Food and Drug Administration (FDA)-approved vaccines or therapeutics available for the prevention or treatment of SARS-CoV-2 infection and/or associated COVID-19 disease, which has triggered a large influx of scientific efforts to develop countermeasures to control SARS-CoV-2 spread. To contribute to these efforts, we have developed an infectious cDNA clone of the SARS-CoV-2 USA-WA1/2020 strain based on the use of a bacterial artificial chromosome (BAC).Recombinant (r)SARS-CoV-2 was readily rescued by transfection of the BAC into Vero E6 cells. Importantly, the BAC-derived rSARS-CoV-2 exhibited growth properties and plaque sizes in cultured cells comparable to those of the SARS-CoV-2 natural isolate. Likewise, rSARS-CoV-2 showed similar levels of replication to that of the natural isolate in nasal turbinates and lungs of infected golden Syrian hamsters. This is, to our knowledge, the first BAC based reverse genetics system for the generation of infectious rSARS-CoV-2 that displays similar features in vivo to that of a natural viral isolate. This SARS-CoV-2 BAC-based reverse genetics will facilitate studies addressing several important questions in the biology of SARS-CoV-2, as well as the identification of antivirals and development of vaccines for the treatment of SARS-CoV-2 infection and associated COVID-19 disease.

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“…The current trends of vaccines against ZIKV mainly concern the pre-membrane and/or envelope (E) proteins as the target epitopes for vaccine candidates due to the natural structure of the virus (Ghaffar et al 2018). The E protein, encompassing the majority of the virion surface, is also the major envelope antigen and is closely related with pathogenicity and infection immunity of ZIKV (Musso and Gubler 2016;Wikan and Smith 2016;Lin et al 2018;Martinez-Sobrido and Almazán 2019). Therefore, the principal aim of the present study was to construct a recombinant ZIKV E protein expressed by baculovirus and assess its immunogenicity and efficacy.…”

Section: Discussionmentioning

confidence: 99%

Baculovirus Surface Display of Zika Virus Envelope Protein Protects against Virus Challenge in Mouse Model

Luo

Miao

et al. 2020

Virol. Sin.

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Zika virus (ZIKV) is emerging as a significant pathogen worldwide and may cause severe neurological disorders such as fetal microcephaly and Guillain-Barre syndrome. No drug or listed vaccines are currently available for preventing ZIKV infection. As a major target of neutralizing, ZIKV envelop (E) protein usually used for vaccine development. Nevertheless, the immunogenicity of ZIKV envelop (E) protein expressed by baculovirus display system has never been assessed. In this study, we reported a new strategy for surface display of ZIKV E protein by a recombinant baculovirus vector derived from Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) and assessed its immunogenicity in mice. We produced recombinant fusion ZIKV E protein linked with signal peptide (SP) and transmembrane domain (TM) of AcMNPV GP64. The results showed that the recombinant protein was easy to produce by baculovirus display system. BALB/c mice immunized with this recombinant E protein developed ZIKV specific serum antibodies. The anti-E protein sera from the mice were able to effectively neutralize ZIKV in vitro. More importantly, AG6 (IFN-a/b and IFN-c receptor deficient) mice immunized with recombinant E protein were protected against lethal ZIKV challenge. Together, these findings demonstrated that the recombinant E protein displayed by baculovirus can be conveniently prepared and displayed good immunogenicity in immunized mice. It is a promising practical approach for prompting the development of vaccine and related immunology research. Keywords Zika virus (ZIKV) Á Envelope protein Á Baculovirus Á Immunogenicity Á Neutralizing antibody Á Viral challenge Dan Luo and Yuanjiu Miao have contributed equally to this work.

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New Advances on Zika Virus Research

Abstract: Zika virus (ZIKV) is an emerging mosquito-borne member of the Flaviviridae family that has historically been known to cause sporadic outbreaks, associated with a mild febrile illness, in Africa and Southeast Asia [...]

Cited by 4 publications

References 35 publications

Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome

Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome

Rescue of SARS-CoV-2 from a single bacterial artificial chromosome

Baculovirus Surface Display of Zika Virus Envelope Protein Protects against Virus Challenge in Mouse Model

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