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  1. Article ; Online: Bacterial Artificial Chromosome-Based Lambda Red Recombination with the I-SceI Homing Endonuclease for Genetic Alteration of MERS-CoV.

    Fehr, Anthony R

    Methods in molecular biology (Clifton, N.J.)

    2020  Volume 2099, Page(s) 53–68

    Abstract: Over the past two decades, several coronavirus (CoV) infectious clones have been engineered, allowing for the manipulation of their large viral genomes (~30 kb) using unique reverse genetic systems. These reverse genetic systems include targeted ... ...

    Abstract Over the past two decades, several coronavirus (CoV) infectious clones have been engineered, allowing for the manipulation of their large viral genomes (~30 kb) using unique reverse genetic systems. These reverse genetic systems include targeted recombination, in vitro ligation, vaccinia virus vectors, and bacterial artificial chromosomes (BACs). Quickly after the identification of Middle East respiratory syndrome-CoV (MERS-CoV), both in vitro ligation and BAC-based reverse genetic technologies were engineered for MERS-CoV to study its basic biological properties, develop live-attenuated vaccines, and test antiviral drugs. Here, I will describe how lambda red recombination can be used with the MERS-CoV BAC to quickly and efficiently introduce virtually any type of genetic modification (point mutations, insertions, deletions) into the MERS-CoV genome and recover recombinant virus.
    MeSH term(s) Bacteriophage lambda/genetics ; Chromosomes, Artificial, Bacterial/genetics ; Coronavirus Infections/prevention & control ; Coronavirus Infections/virology ; Deoxyribonuclease I/genetics ; Deoxyribonuclease I/metabolism ; Genetic Engineering ; Genome, Viral/genetics ; Homologous Recombination ; Humans ; Middle East Respiratory Syndrome Coronavirus/genetics ; Middle East Respiratory Syndrome Coronavirus/immunology ; Mutation ; Vaccines, Attenuated/genetics ; Vaccinia virus/genetics ; Viral Vaccines/genetics
    Chemical Substances Vaccines, Attenuated ; Viral Vaccines ; Deoxyribonuclease I (EC 3.1.21.1)
    Keywords covid19
    Language English
    Publishing date 2020-01-09
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-0211-9_5
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: PARP14: A key ADP-ribosylating protein in host-virus interactions?

    Parthasarathy, Srivatsan / Fehr, Anthony R

    PLoS pathogens

    2022  Volume 18, Issue 6, Page(s) e1010535

    MeSH term(s) ADP Ribose Transferases/metabolism ; Adenosine Diphosphate ; Host Microbial Interactions ; Poly(ADP-ribose) Polymerases/metabolism
    Chemical Substances Adenosine Diphosphate (61D2G4IYVH) ; ADP Ribose Transferases (EC 2.4.2.-) ; Poly(ADP-ribose) Polymerases (EC 2.4.2.30)
    Language English
    Publishing date 2022-06-09
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2205412-1
    ISSN 1553-7374 ; 1553-7374
    ISSN (online) 1553-7374
    ISSN 1553-7374
    DOI 10.1371/journal.ppat.1010535
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: PARP14

    Srivatsan Parthasarathy / Anthony R Fehr

    PLoS Pathogens, Vol 18, Iss 6, p e

    A key ADP-ribosylating protein in host-virus interactions?

    2022  Volume 1010535

    Keywords Immunologic diseases. Allergy ; RC581-607 ; Biology (General) ; QH301-705.5
    Language English
    Publishing date 2022-06-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article: An Update on the Current State of SARS-CoV-2 Mac1 Inhibitors.

    O'Connor, Joseph J / Ferraris, Dana / Fehr, Anthony R

    Pathogens (Basel, Switzerland)

    2023  Volume 12, Issue 10

    Abstract: Non-structural protein 3 (nsp3) from all coronaviruses (CoVs) contains a conserved macrodomain, known as Mac1, that has been proposed as a potential therapeutic target for CoVs due to its critical role in viral pathogenesis. Mac1 is an ADP-ribose binding ...

    Abstract Non-structural protein 3 (nsp3) from all coronaviruses (CoVs) contains a conserved macrodomain, known as Mac1, that has been proposed as a potential therapeutic target for CoVs due to its critical role in viral pathogenesis. Mac1 is an ADP-ribose binding protein and ADP-ribosylhydrolase that promotes replication and blocks IFN responses, though the precise mechanisms it uses to carry out these functions remain unknown. Over the past 3 years following the onset of COVID-19, several groups have used high-throughput screening with multiple assays and chemical modifications to create unique chemical inhibitors of the SARS-CoV-2 Mac1 protein. Here, we summarize the current efforts to identify selective and potent inhibitors of SARS-CoV-2 Mac1.
    Language English
    Publishing date 2023-10-07
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2695572-6
    ISSN 2076-0817
    ISSN 2076-0817
    DOI 10.3390/pathogens12101221
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Bacterial Artificial Chromosome-Based Lambda Red Recombination with the I-SceI Homing Endonuclease for Genetic Alteration of MERS-CoV

    Fehr, Anthony R

    Methods Mol Biol

    Abstract: Over the past two decades, several coronavirus (CoV) infectious clones have been engineered, allowing for the manipulation of their large viral genomes (~30 kb) using unique reverse genetic systems. These reverse genetic systems include targeted ... ...

    Abstract Over the past two decades, several coronavirus (CoV) infectious clones have been engineered, allowing for the manipulation of their large viral genomes (~30 kb) using unique reverse genetic systems. These reverse genetic systems include targeted recombination, in vitro ligation, vaccinia virus vectors, and bacterial artificial chromosomes (BACs). Quickly after the identification of Middle East respiratory syndrome-CoV (MERS-CoV), both in vitro ligation and BAC-based reverse genetic technologies were engineered for MERS-CoV to study its basic biological properties, develop live-attenuated vaccines, and test antiviral drugs. Here, I will describe how lambda red recombination can be used with the MERS-CoV BAC to quickly and efficiently introduce virtually any type of genetic modification (point mutations, insertions, deletions) into the MERS-CoV genome and recover recombinant virus.
    Keywords covid19
    Publisher WHO
    Document type Article
    Note WHO #Covidence: #710836
    Database COVID19

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  6. Article: The Conserved Macrodomain Is a Potential Therapeutic Target for Coronaviruses and Alphaviruses.

    Leung, Anthony K L / Griffin, Diane E / Bosch, Jürgen / Fehr, Anthony R

    Pathogens (Basel, Switzerland)

    2022  Volume 11, Issue 1

    Abstract: Emerging and re-emerging viral diseases pose continuous public health threats, and effective control requires a combination of non-pharmacologic interventions, treatment with antivirals, and prevention with vaccines. The COVID-19 pandemic has ... ...

    Abstract Emerging and re-emerging viral diseases pose continuous public health threats, and effective control requires a combination of non-pharmacologic interventions, treatment with antivirals, and prevention with vaccines. The COVID-19 pandemic has demonstrated that the world was least prepared to provide effective treatments. This lack of preparedness has been due, in large part, to a lack of investment in developing a diverse portfolio of antiviral agents, particularly those ready to combat viruses of pandemic potential. Here, we focus on a drug target called macrodomain that is critical for the replication and pathogenesis of alphaviruses and coronaviruses. Some mutations in alphavirus and coronaviral macrodomains are not tolerated for virus replication. In addition, the coronavirus macrodomain suppresses host interferon responses. Therefore, macrodomain inhibitors have the potential to block virus replication and restore the host's protective interferon response. Viral macrodomains offer an attractive antiviral target for developing direct acting antivirals because they are highly conserved and have a structurally well-defined (druggable) binding pocket. Given that this target is distinct from the existing RNA polymerase and protease targets, a macrodomain inhibitor may complement current approaches, pre-empt the threat of resistance and offer opportunities to develop combination therapies for combating COVID-19 and future viral threats.
    Language English
    Publishing date 2022-01-14
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2695572-6
    ISSN 2076-0817
    ISSN 2076-0817
    DOI 10.3390/pathogens11010094
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: The Conserved Macrodomain Is a Potential Therapeutic Target for Coronaviruses and Alphaviruses

    Anthony K. L. Leung / Diane E. Griffin / Jürgen Bosch / Anthony R. Fehr

    Pathogens, Vol 11, Iss 94, p

    2022  Volume 94

    Abstract: Emerging and re-emerging viral diseases pose continuous public health threats, and effective control requires a combination of non-pharmacologic interventions, treatment with antivirals, and prevention with vaccines. The COVID-19 pandemic has ... ...

    Abstract Emerging and re-emerging viral diseases pose continuous public health threats, and effective control requires a combination of non-pharmacologic interventions, treatment with antivirals, and prevention with vaccines. The COVID-19 pandemic has demonstrated that the world was least prepared to provide effective treatments. This lack of preparedness has been due, in large part, to a lack of investment in developing a diverse portfolio of antiviral agents, particularly those ready to combat viruses of pandemic potential. Here, we focus on a drug target called macrodomain that is critical for the replication and pathogenesis of alphaviruses and coronaviruses. Some mutations in alphavirus and coronaviral macrodomains are not tolerated for virus replication. In addition, the coronavirus macrodomain suppresses host interferon responses. Therefore, macrodomain inhibitors have the potential to block virus replication and restore the host’s protective interferon response. Viral macrodomains offer an attractive antiviral target for developing direct acting antivirals because they are highly conserved and have a structurally well-defined (druggable) binding pocket. Given that this target is distinct from the existing RNA polymerase and protease targets, a macrodomain inhibitor may complement current approaches, pre-empt the threat of resistance and offer opportunities to develop combination therapies for combating COVID-19 and future viral threats.
    Keywords coronavirus ; alphavirus ; SARS-CoV-2 ; macrodomain ; ADP-ribosylation ; ADP-ribosylhydrolase ; Medicine ; R
    Subject code 570
    Language English
    Publishing date 2022-01-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  8. Article: Two Commercially Available Blood-Stabilization Reagents Serve as Potent Inactivators of Coronaviruses.

    O'Connor, Joseph J / Voth, Lynden / Athmer, Jeremiah / George, Nicholas M / Connelly, Christopher M / Fehr, Anthony R

    Pathogens (Basel, Switzerland)

    2023  Volume 12, Issue 9

    Abstract: The continued circulation of SARS-CoV-2 and the increasing frequency of coronavirus (CoV) outbreaks over the decades demonstrates the enduring threat that the CoV family poses. There remains a significant need to develop tools to monitor and prevent the ... ...

    Abstract The continued circulation of SARS-CoV-2 and the increasing frequency of coronavirus (CoV) outbreaks over the decades demonstrates the enduring threat that the CoV family poses. There remains a significant need to develop tools to monitor and prevent the spread of these viruses. We tested blood-stabilization reagents from two commercially available blood collection tubes (BCTs) for their ability to inactivate three different coronaviruses (MHV, OC-43, and SARS-CoV-2) and stabilize their RNA. Both Cell-Free DNA BCT
    Language English
    Publishing date 2023-08-25
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2695572-6
    ISSN 2076-0817
    ISSN 2076-0817
    DOI 10.3390/pathogens12091082
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  9. Article: The Conserved Macrodomain Is a Potential Therapeutic Target for Coronaviruses and Alphaviruses

    Leung, Anthony K. L. / Griffin, Diane E. / Bosch, Jürgen / Fehr, Anthony R.

    Pathogens. 2022 Jan. 14, v. 11, no. 1

    2022  

    Abstract: Emerging and re-emerging viral diseases pose continuous public health threats, and effective control requires a combination of non-pharmacologic interventions, treatment with antivirals, and prevention with vaccines. The COVID-19 pandemic has ... ...

    Abstract Emerging and re-emerging viral diseases pose continuous public health threats, and effective control requires a combination of non-pharmacologic interventions, treatment with antivirals, and prevention with vaccines. The COVID-19 pandemic has demonstrated that the world was least prepared to provide effective treatments. This lack of preparedness has been due, in large part, to a lack of investment in developing a diverse portfolio of antiviral agents, particularly those ready to combat viruses of pandemic potential. Here, we focus on a drug target called macrodomain that is critical for the replication and pathogenesis of alphaviruses and coronaviruses. Some mutations in alphavirus and coronaviral macrodomains are not tolerated for virus replication. In addition, the coronavirus macrodomain suppresses host interferon responses. Therefore, macrodomain inhibitors have the potential to block virus replication and restore the host’s protective interferon response. Viral macrodomains offer an attractive antiviral target for developing direct acting antivirals because they are highly conserved and have a structurally well-defined (druggable) binding pocket. Given that this target is distinct from the existing RNA polymerase and protease targets, a macrodomain inhibitor may complement current approaches, pre-empt the threat of resistance and offer opportunities to develop combination therapies for combating COVID-19 and future viral threats.
    Keywords Alphavirus ; COVID-19 infection ; DNA-directed RNA polymerase ; Orthocoronavirinae ; antiviral agents ; interferons ; pandemic ; pathogenesis ; proteinases ; public health ; therapeutics ; virus replication
    Language English
    Dates of publication 2022-0114
    Publishing place Multidisciplinary Digital Publishing Institute
    Document type Article
    ZDB-ID 2695572-6
    ISSN 2076-0817
    ISSN 2076-0817
    DOI 10.3390/pathogens11010094
    Database NAL-Catalogue (AGRICOLA)

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  10. Article ; Online: The Viral Macrodomain Counters Host Antiviral ADP-Ribosylation.

    Alhammad, Yousef M O / Fehr, Anthony R

    Viruses

    2020  Volume 12, Issue 4

    Abstract: Macrodomains, enzymes that remove ADP-ribose from proteins, are encoded by several families of RNA viruses and have recently been shown to counter innate immune responses to virus infection. ADP-ribose is covalently attached to target proteins by poly- ... ...

    Abstract Macrodomains, enzymes that remove ADP-ribose from proteins, are encoded by several families of RNA viruses and have recently been shown to counter innate immune responses to virus infection. ADP-ribose is covalently attached to target proteins by poly-ADP-ribose polymerases (PARPs), using nicotinamide adenine dinucleotide (NAD+) as a substrate. This modification can have a wide variety of effects on proteins including alteration of enzyme activity, protein-protein interactions, and protein stability. Several PARPs are induced by interferon (IFN) and are known to have antiviral properties, implicating ADP-ribosylation in the host defense response and suggesting that viral macrodomains may counter this response. Recent studies have demonstrated that viral macrodomains do counter the innate immune response by interfering with PARP-mediated antiviral defenses, stress granule formation, and pro-inflammatory cytokine production. Here, we will describe the known functions of the viral macrodomains and review recent literature demonstrating their roles in countering PARP-mediated antiviral responses.
    MeSH term(s) ADP-Ribosylation/immunology ; Adenosine Diphosphate Ribose/metabolism ; Cytoplasmic Granules/immunology ; Cytoplasmic Granules/virology ; Humans ; Interferons/immunology ; Mutation ; Poly(ADP-ribose) Polymerases/immunology ; Protein Domains ; RNA Virus Infections/immunology ; RNA Virus Infections/metabolism ; RNA Virus Infections/virology ; RNA Viruses/classification ; RNA Viruses/genetics ; RNA Viruses/immunology ; RNA Viruses/metabolism ; Viral Nonstructural Proteins/chemistry ; Viral Nonstructural Proteins/genetics ; Viral Nonstructural Proteins/immunology ; Viral Nonstructural Proteins/metabolism ; Virus Replication
    Chemical Substances Viral Nonstructural Proteins ; Adenosine Diphosphate Ribose (20762-30-5) ; Interferons (9008-11-1) ; Poly(ADP-ribose) Polymerases (EC 2.4.2.30)
    Keywords covid19
    Language English
    Publishing date 2020-03-31
    Publishing country Switzerland
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Review
    ZDB-ID 2516098-9
    ISSN 1999-4915 ; 1999-4915
    ISSN (online) 1999-4915
    ISSN 1999-4915
    DOI 10.3390/v12040384
    Database MEDical Literature Analysis and Retrieval System OnLINE

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