======================
   WUHAN
 ======================

JOURNAL OF VIROLOGY, Feb. 2008, p. 1899–1907
0022-538X/08/$08.00ϩ0 doi:10.1128/JVI.01085-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.
Vol. 82, No. 4

Difference in Receptor Usage between Severe Acute Respiratory
Syndrome (SARS) Coronavirus and SARS-Like
Coronavirus of Bat Originᰔ

Wuze Ren,1† Xiuxia Qu,2† Wendong Li,1‡ Zhenggang Han,1 Meng Yu,3 Peng Zhou,1 Shu-Yi Zhang,4
Lin-Fa Wang,3* Hongkui Deng,2 and Zhengli Shi1*
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China1; Key Laboratory of
Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, China2;
CSIRO Livestock Industries, Australian Animal Health Laboratory and Australian Biosecurity Cooperative Research Center for
Emerging Infectious Diseases, Geelong, Australia3; and School of Life Science, East China Normal University, Shanghai, China 4
Received 20 May 2007/Accepted 15 November 2007


  ======================
    BARIC:
  ======================

-- Baric on Coronavirus as early as 1988, 1990, 1999, 2000 (examples):

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    Baric R. S., Schaad M. C., Stohlman S.
Establishing a genetic recombination map for the murine coronavirus strain A59 complementation groups
Virology 177 1990 646 656

    Baric R. S., Sullivan E., Hensley L., Yount B., Chen W.
Persistent infection promotes cross-species transmissibility of mouse hepatitis virus.
J. Virol. 71 1999 638 649

    Baric R. S., Yount B.
Subgenomic negative-strand function during mouse hepatitis virus infection.
J. Virol. 74 2000 4039 4046


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-- and on coronavirus chimera construction (2000):

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DOI: 10.1128/JVI.74.22.10600-10611.2000

REPLICATION

Strategy for Systematic Assembly of Large RNA and DNA Genomes:
Transmissible Gastroenteritis Virus Model

Boyd Yount, Kristopher M. Curtis, Ralph S. Baric

ABSTRACT

A systed was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in
nature. The ∼30-kb genome, coupled with regions of genomic instability, has hindered the development of a full-length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine.

ACKNOWLEDGMENTS
[...]
This work was supported by a research grant from the National Institutes of Health (AI 23946).

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Baric/NCU 2008:

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Proc Natl Acad Sci U S A. 2008 Dec 16; 105(50): 19944–19949.
Published online 2008 Nov 26. doi: 10.1073/pnas.0808116105
PMCID: PMC2588415
PMID: 19036930

Microbiology

Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice

Michelle M. Becker/a,/1, Rachel L. Graham/b,/1, Eric F. Donaldson/b. Barry Rockx/b, Amy C. Sims/b,/c, Timothy Sheahan/b, Raymond J. Pickles/d,/e, Davide Corti/f, Robert E. Johnston/c, Ralph S. Baric/b,/c,/d,/2 and Mark R. Denisona/g,/2

Author information
Departments of /a/Pediatrics and
/g/Microbiology and Immunology, Vanderbilt University, Nashville, TN 37232;
Departments of /b/Epidemiology and
/d/Microbiology and Immunology,
/e/Cystic Fibrosis/Pulmonary Research and Treatment Center and Department of Medicine, and
/c/Carolina Vaccine Institute, University of North Carolina, Chapel Hill, NC 27599; and
/f/Institute for Research in Biomedicine, CH-6500 Bellinzona, Switzerland

/2/To whom correspondence may be addressed. E-mail: ude.cnu.liame@cirabr or ude.tlibrednav@nosined.kram
Edited by Peter Palese, Mount Sinai School of Medicine, New York, NY, and approved October 14, 2008

Abstract
[...]  To test a possible route of emergence from the noncultivable Bat-SCoV to human SARS-CoV, we designed a consensus Bat-SCoV genome and replaced the Bat-SCoV Spike receptor-binding domain (RBD) with the SARS-CoV RBD (Bat-SRBD). Bat-SRBD was infectious in cell culture and in mice and was efficiently neutralized by antibodies specific for both bat and human CoV Spike proteins. Rational design, synthesis, and recovery of hypothetical recombinant viruses can be used to investigate mechanisms of transspecies movement of zoonoses and has great potential to aid in rapid public health responses to known or predicted emerging microbial threats.


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 -   Baric-Shi:   //Chapel-Hill-2015.htm

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REM: "nature nature medicine letters article"
    PMID: 26552008
    PMCID: PMC4797993
    DOI: 10.1038/nm.3985

Nature Medicine
 2015 Dec;21(12):1508-13.
doi: 10.1038/nm.3985. Epub 2015 Nov 9.

A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence

Vineet D Menachery  1 , Boyd L Yount Jr  1 , Kari Debbink  1   2 , Sudhakar Agnihothram  3 , Lisa E Gralinski  1 , Jessica A Plante  1 , Rachel L Graham  1 , Trevor Scobey  1 , Xing-Yi Ge  4 , Eric F Donaldson  1 , Scott H Randell  5   6 , Antonio Lanzavecchia  7 , Wayne A Marasco  8   9 , Zhengli-Li Shi  4 , Ralph S Baric  1   2

Affiliations

    1 Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
    2 Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
    3 National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arkansas, USA.
    4 Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
    5 Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
    6 Cystic Fibrosis Center, Marsico Lung Institute, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
    7 Institute for Research in Biomedicine, Bellinzona Institute of Microbiology, Zurich, Switzerland.
    8 Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA.
    9 Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.


AUTHOR CONTRIBUTIONS
...  Z.-L.S. [Shi] provided SHC014 spike sequences and
plasmids. R.S.B. [Baric] designed experiments and wrote manuscript.

[...]
Here we examine the disease potential of a SARS-like virus, SHC014-CoV,
which is currently circulating in Chinese horseshoe bat populations1.
Using the SARS-CoV reverse genetics system2, we generated and
characterized a chimeric virus expressing the spike of bat
coronavirus SHC014 in a mouse-adapted SARS-CoV backbone.
The results indicate that group 2b viruses encoding the SHC014 spike
in a wild-type backbone can [...] replicate efficiently in primary human
airway cells and achieve in vitro titers equivalent to epidemic strains of
SARS-CoV. Additionally, in vivo experiments demonstrate replication of
the chimeric virus in mouse lung with notable pathogenesis.
[..]

[Chimera construction:]
Wild-type SARS-CoV (Urbani), mouse-adapted SARS-CoV (MA15) and
chimeric SARS-like CoVs were cultured on Vero E6 cells (obtained from
United States Army Medical arch Institute of Infectious Diseases),{...]
DBT cells (Baric laboratory, source unknown) expressing ACE2 orthologs have
been previously described for both human and civet; bat Ace2 sequence was
based on that from Rhinolophus leschenaulti, and DBT cells expressing bat
Ace2 were established as described previously8. Pseudotyping experiments
were similar to those using an HIV-based pseudovirus, prepared as previously
described10, and examined on HeLa cells (Wuhan Institute of Virology) that
expressed ACE2 orthologs. HeLa cells were grown in minimal essential medium
(MEM) (Gibco, CA) supplemented with 10% FCS (Gibco, CA) as previously
described24.

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 - in kind of a "current overview" Textbook

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Current Opinion in Virology
Volume 23, April 2017, Pages 1-7

https://doi.org/10.1016/j.coviro.2017.01.002

Jumping species—a mechanism for coronavirus persistence and survival

Vineet D  Menachery Rachel L Graham  Ralph S Baric

    Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States
Available online 31 March 2017.

{from part:] Keying in: spike drives emergence

[...]
With binding required for infection, mutations within S1, and most notably, the receptor-binding domain (RBD), have been thought to be critical for CoV emergence [30]. Using chimeric viruses employing civet, early, and middle-phase spike proteins demonstrated viability for the closely related strains in human cells [31, 32]. However, for some strains, such as SZ16 and bat-derived HKU3-CoV, the closest known SARS-CoV progenitors at the time, progeny virions were not recoverable in Vero or primary human airway epithelial cells, despite evidence of RNA replication [30, 32]. To overcome this barrier, single humanizing mutation K479N was introduced into SZ16 and a chimeric HKU3 virus containing the RBD of SARS-CoV was designed and permitted replication, likely due to its capacity to bind the human ACE2 receptor [30, 31]. A similar approach was used with group 2C CoV HKU5; substitution of the entire ecto-domain from SARS-CoV spike resulted in an HKU5 virus that was able to infect human cells [33]. Together, the data argue that the ability of the spike to bind receptor is required for viability in novel hosts.

[ References: ]
31
    T. Sheahan, B. Rockx, E. Donaldson, D. Corti, R. Baric
    Pathways of cross-species transmission of synthetically reconstructed zoonotic severe acute respiratory syndrome coronavirus
    J Virol, 82 (2008), pp. 8721-8732
    View Record in ScopusGoogle Scholar
32
    T. Sheahan, B. Rockx, E. Donaldson, A. Sims, R. Pickles, D. Corti, R. Baric
    Mechanisms of zoonotic severe acute respiratory syndrome coronavirus host range expansion in human airway epithelium
    J Virol, 82 (2008), pp. 2274-2285
    View Record in ScopusGoogle Scholar
33
    S. Agnihothram, B.L. Yount Jr., E.F. Donaldson, J. Huynh, V.D. Menachery, L.E. Gralinski, R.L. Graham, M.M. Becker, S. Tomar, T.D. Scobey, et al.
    A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant
    MBio, 5 (2014), pp. e00047-00014
    Google Scholar
34
    V.D. Menachery, B.L. Yount Jr., K. Debbink, S. Agnihothram, L.E. Gralinski, J.A. Plante, R.L. Graham, T. Scobey, X.Y. Ge, E.F. Donaldson, et al.
    A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence
    Nat Med, 21 (2015), pp. 1508-1513
    CrossRefView Record in ScopusGoogle Scholar
35
    V.D. Menachery, B.L. Yount Jr., A.C. Sims, K. Debbink, S.S. Agnihothram, L.E. Gralinski, R.L. Graham, T. Scobey, J.A. Plante, S.R. Royal, et al.
    SARS-like WIV1-CoV poised for human emergence
    Proc Natl Acad Sci U S A, 113 (2016), pp. 3048-3053
    CrossRefView Record in ScopusGoogle Scholar


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  -- in defending GoF, 2018; //Baric-4GoF-2018.html

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Published ... Future Virol. 2018 Mar; 13(3): 143–146.
Published online 2018 Feb 21. doi: 10.2217/fvl-2017-0143
PMCID: PMC6289267
NIHMSID: NIHMS973008
PMID: 30546388

Is regulation preventing the development of therapeutics that may prevent future coronavirus pandemics?

Timothy P Sheahan*,1 and Ralph S Baric1

... To this end, we have constructed recombinant virus from infectious cDNA clones for multiple human and zoonotic CoV [9,1115].

... [P]rior to the ‘pause’, we reconstructed SARS-like (i.e., Bat CoV HKU3) and MERS-like (Bat CoV HKU5) bat CoV that could replicate in mammalian cell lines but could not propagate suggesting receptor and virus spike glycoprotein incompatibilities [15,18].

... This analysis would not have been possible without passage of CoV to increase pathogenicity, the shifting of zoonotic CoV host range to infect human cells, and the reconstruction of ‘prepandemic’ SARS-like bat CoV poised for emergence (i.e., WIV1, SCH014).

... This analysis [in view of a MERS drug] would not have been possible without passage of CoV to increase pathogenicity, the shifting of zoonotic CoV host range to infect human cells, and the reconstruction of ‘prepandemic’ SARS-like bat CoV poised for emergence [i.e., WIV1, SCH014].

--> [9] [SELF-REFERENCIAL:
PNAS March 15, 2016 113 (11) 3048-3053; first published March 14, 2016; https://doi.org/10.1073/pnas.1517719113
Baric, Sheahan, e.a (18 ! authors, all UNC)
SARS-like WIV1-CoV poised for human emergence
... Synthetic construction of chimeric mutant and full-length WIV1-CoV were approved by the UNC Institutional Biosafety Committee and the Dual Use Research of Concern Committee.
... We thank Dr. Zhengli-Li Shi of the Wuhan Institute of Virology for access to bat CoV sequences and plasmid of WIV1-CoV spike protein.

--> [11] Schon VOR SARS, bricolage mit CoV [2002]:

Systematic assembly of a full-length infectious cDNA of mouse hepatitis virus strain A59
B Yount, MR Denison, SR Weiss, RS Baric
- Journal of virology, 2002 - Am Soc Microbiol

A novel method was developed to assemble a full-length infectious cDNA of the group II
coronavirus mouse hepatitis virus strain A59 (MHV-A59). Seven contiguous cDNA clones
that spanned the 31.5-kb MHV genome were isolated. The ends of the cDNAs were
engineered with unique junctions and assembled with only the adjacent cDNA subclones

--> [15]
A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence
VD Menachery, BL Yount, K Debbink, S Agnihothram… [e.a. Baric, all NUC]
 - Nature medicine, 2015 -

... Here we examine the disease potential
of a SARS-like virus, SHC014-CoV, which is currently circulating in Chinese horseshoe bat
populations 1. Using the SARS-CoV reverse genetics system 2, we generated and
characterized a chimeric virus expressing the spike of bat coronavirus SHC014 in a mouse …

--> [18]
Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice
Micelle M. Becker, Rachel L. Graham, Eric F. Donaldson, Barry Rockx, Amy C. Sims, Timothy Sheahan, Raymond J. Pickles, Davide Corti, Robert E. Johnston, Ralph S. Baric, and Mark R. Denison
PNAS December 16, 2008 105 (50) 19944-19949; https://doi.org/10.1073/pnas.0808116105


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  -- Baric and Remdesivir/Gilead:

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https://www.technologynetworks.com/genomics/news/what-the-d614g-mutation-means-for-sars-cov-2-vaccines-342786:

Baric has studied coronaviruses for more than three decades and was integral in the development of remdesivir, the first FDA-approved treatment for COVID-19.

REM:
-- Gilead founder: Donald Rumsfeld:
-- NIAID support for remdesivir:

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