Political Hit
The Iran Attack on Israel
was planned through Centcom
and the Swiss, and 6 nations.
Purpose: prevent a bigger war
and make Sleepy Joe seem
like he was in charge ...
Electioneering .
Then there's the continued
COVID Lab Origin denials.
2008 proof
Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice
Michelle M. Becker, Rachel L. Graham, Eric F. Donaldson, +7, and Mark R. Denison rbaric@email.unc.edu
October 14, 2008
Here, we report the design, synthesis, and recovery of the largest synthetic replicating life form, a 29.7-kb bat severe acute respiratory syndrome (SARS)-like coronavirus (Bat-SCoV), a likely progenitor to the SARS-CoV epidemic.
Only 2500 people looked
at THIS in the past YEAR !
PNAS Logo
RESEARCH ARTICLE
OPEN ACCESS
SHARE ON
Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice
Michelle M. Becker, Rachel L. Graham, Eric F. Donaldson, +7, and Mark R. Denison rbaric@email.unc.eduAuthors Info & Affiliations
Edited by Peter Palese, Mount Sinai School of Medicine, New York, NY, and approved October 14, 2008
December 16, 2008
105 (50) 19944-19949
https://doi.org/10.1073/pnas.0808116105
79,675
180
Metrics
Total views
79,675
Last 12 Months
2,570
Total citations
180
Last 12 Months
6
Vol. 105 | No. 50
Abstract
Data Availability
Acknowledgments.
Supporting Information
References
Information & Authors
Metrics & Citations
View Options
References
Share
Abstract
Defining prospective pathways by which zoonoses evolve and emerge as human pathogens is critical for anticipating and controlling both natural and deliberate pandemics. However, predicting tenable pathways of animal-to-human movement has been hindered by challenges in identifying reservoir species, cultivating zoonotic organisms in culture, and isolating full-length genomes for cloning and genetic studies. The ability to design and recover pathogens reconstituted from synthesized cDNAs has the potential to overcome these obstacles by allowing studies of replication and pathogenesis without identification of reservoir species or cultivation of primary isolates. Here, we report the design, synthesis, and recovery of the largest synthetic replicating life form, a 29.7-kb bat severe acute respiratory syndrome (SARS)-like coronavirus (Bat-SCoV), a likely progenitor to the SARS-CoV epidemic. 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.
Continue Reading
Data Availability
Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. FJ211859 and FJ211860).
Acknowledgments.
We thank XiaoTao Lu and Sunny Lee for technical assistance, Susan Burkett for maintenance of the HAE cultures, Perry Myrick for immunofluorescence assays, and the University of North Carolina Cystic Fibrosis Tissue Culture Core for HAE cells. M.M.B., R.L.G., R.S.B., and M.R.D. are supported by the National Institute of Allergy and Infectious Diseases Public Health Service Award P01 AI59943. Additional support was provided by Public Health Service Award CA68485 to the Vanderbilt University DNA Sequencing Shared Resource of the Vanderbilt–Ingram Cancer Center. The Baric laboratory is supported by the Gillings Innovation Fund.
Supporting Information
Supporting Information (PDF)
Supporting Information
DOWNLOAD
3.19 MB
References
1
RA Weiss, AJ McMichael, Social and environmental risk factors in the emergence of infectious diseases. Nat Med 10, S70–76 (2004).
Crossref
PubMed
Google Scholar
2
ME Woolhouse, S Gowtage-Sequeria, Host range and emerging and reemerging pathogens. Emerg Infect Dis 11, 1842–1847 (2005).
Crossref
PubMed
Google Scholar
3
R Webby, E Hoffmann, R Webster, Molecular constraints to interspecies transmission of viral pathogens. Nat Med 10, S77–81 (2004).
Crossref
PubMed
Google Scholar
4
C Drosten, et al., Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med 348, 1967–1976 (2003).
Crossref
PubMed
Google Scholar
5
TG Ksiazek, et al., A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med 348, 1953–1966 (2003).
Crossref
PubMed
Google Scholar
6
JS Peiris, et al., Coronavirus as a possible cause of severe acute respiratory syndrome. Lancet 361, 1319–1325 (2003).
Crossref
PubMed
Google Scholar
7
C Tu, et al., Antibodies to SARS coronavirus in civets. Emerg Infect Dis 10, 2244–2248 (2004).
Crossref
PubMed
Google Scholar
8
B Kan, et al., Molecular evolution analysis and geographic investigation of severe acute respiratory syndrome coronavirus-like virus in palm civets at an animal market and on farms. J Virol 79, 11892–11900 (2005).
Crossref
PubMed
Google Scholar
9
Y Guan, et al., Isolation and characterization of viruses related to the SARS coronavirus from animals in southern China. Science 302, 276–278 (2003).
Crossref
PubMed
Google Scholar
10
W Li, et al., Bats are natural reservoirs of SARS-like coronaviruses. Science 310, 676–679 (2005).
Crossref
PubMed
Google Scholar
11
Z Shi, Z Hu, A review of studies on animal reservoirs of the SARS coronavirus. Virus Res 133, 74–87 (2008).
Crossref
PubMed
Google Scholar
12
SK Lau, et al., Severe acute respiratory syndrome coronavirus-like virus in Chinese horseshoe bats. Proc Natl Acad Sci USA 102, 14040–14045 (2005).
Crossref
PubMed
Google Scholar
13
SR Dominguez, TJ O'Shea, LM Oko, KV Holmes, Detection of group 1 coronaviruses in bats in North America. Emerg Infect Dis 13, 1295–1300 (2007).
Crossref
PubMed
Google Scholar
14
K Halpin, et al., Emerging viruses: Coming in on a wrinkled wing and a prayer. Clin Infect Dis 44, 711–717 (2007).
Crossref
PubMed
Google Scholar
15
CH Calisher, et al., Bats: Important reservoir hosts of emerging viruses. Clin Microbiol Rev 19, 531–545 (2006).
Crossref
PubMed
Google Scholar
16
EC Holmes, A Rambaut, Viral evolution and the emergence of SARS coronavirus. Philos Trans R Soc Lond B Biol Sci 359, 1059–1065 (2004).
Crossref
PubMed
Google Scholar
17
MM Lai, D Cavanagh, The molecular biology of coronaviruses. Adv Virus Res 48, 1–100 (1997).
Crossref
PubMed
Google Scholar
18
J Stavrinides, DS Guttman, Mosaic evolution of the severe acute respiratory syndrome coronavirus. J Virol 78, 76–82 (2004).
Crossref
PubMed
Google Scholar
19
W Li, et al., Animal origins of the severe acute respiratory syndrome coronavirus: Insight from ACE2-S-protein interactions. J Virol 80, 4211–4219 (2006).
Crossref
PubMed
Google Scholar
20
W Li, et al., The S proteins of human coronavirus NL63 and severe acute respiratory syndrome coronavirus bind overlapping regions of ACE2. Virology 367, 367–374 (2007).
Crossref
PubMed
Google Scholar
21
W Li, et al., Receptor and viral determinants of SARS-coronavirus adaptation to human ACE2. EMBO J 24, 1634–1643 (2005).
Crossref
PubMed
Google Scholar
22
T Sheahan, et al., Mechanisms of zoonotic severe acute respiratory syndrome coronavirus host range expansion in human airway epithelium. J Virol 82, 2274–2285 (2008).
Crossref
PubMed
Google Scholar
23
KD Pruitt, T Tatusova, DR Maglott, NCBI Reference Sequence project: Update and current status. Nucleic Acids Res 31, 34–37 (2003).
Crossref
PubMed
Google Scholar
24
B Yount, et al., Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus. Proc Natl Acad Sci USA 100, 12995–13000 (2003).
Crossref
PubMed
Google Scholar
25
L Kuo, et al., Retargeting of coronavirus by substitution of the spike glycoprotein ectodomain: Crossing the host cell species barrier. J Virol 74, 1393–1406 (2000).
Crossref
PubMed
Google Scholar
26
BJ Haijema, H Volders, PJ Rottier, Switching species tropism: An effective way to manipulate the feline coronavirus genome. J Virol 77, 4528–4538 (2003).
Crossref
PubMed
Google Scholar
27
S Chakraborti, P Prabakaran, X Xiao, DS Dimitrov, The SARS coronavirus S glycoprotein receptor binding domain: Fine mapping and functional characterization. Virol J 2, 73 (2005).
Crossref
PubMed
Google Scholar
28
SK Wong, et al., A 193-amino acid fragment of the SARS coronavirus S protein efficiently binds angiotensin-converting enzyme 2. J Biol Chem 279, 3197–3201 (2004).
Crossref
PubMed
Google Scholar
29
F Li, W Li, M Farzan, SC Harrison, Structure of SARS coronavirus spike receptor-binding domain complexed with receptor. Science 309, 1864–1868 (2005).
Crossref
PubMed
Google Scholar
30
E Prentice, et al., Identification and characterization of severe acute respiratory syndrome coronavirus replicase proteins. J Virol 78, 9977–9986 (2004).
Crossref
PubMed
Google Scholar
31
B Rockx, et al., Structural basis for potent cross-neutralizing human monoclonal antibody protection against lethal human and zoonotic severe acute respiratory syndrome coronavirus challenge. J Virol 82, 3220–3235 (2008).
Crossref
PubMed
Google Scholar
32
B Rockx, et al., Synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice. J Virol 81, 7410–7423 (2007).
Crossref
PubMed
Google Scholar
33
AC Sims, et al., Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: Role of ciliated cells in viral spread in the conducting airways of the lungs. J Virol 79, 15511–15524 (2005).
Crossref
PubMed
Google Scholar
34
K Subbarao, A Roberts, Is there an ideal animal model for SARS? Trends Microbiol 14, 299–303 (2006).
Crossref
PubMed
Google Scholar
35
A Roberts, et al., Animal models and vaccines for SARS-CoV infection. Virus Res 133, 20–32 (2008).
Crossref
PubMed
Google Scholar
36
A Roberts, et al., A mouse-adapted SARS-coronavirus causes disease and mortality in BALB/c mice. PLoS Pathog 3, e5 (2007).
Crossref
PubMed
Google Scholar
37
HO Smith, CA Hutchison, C Pfannkoch, JC Venter, Generating a synthetic genome by whole genome assembly: PhiX174 bacteriophage from synthetic oligonucleotides. Proc Natl Acad Sci USA 100, 15440–15445 (2003).
Crossref
PubMed
Google Scholar
38
J Cello, AV Paul, E Wimmer, Chemical synthesis of poliovirus cDNA: Generation of infectious virus in the absence of natural template. Science 297, 1016–1018 (2002).
Crossref
PubMed
Google Scholar
39
TM Tumpey, et al., Characterization of the reconstructed 1918 Spanish influenza pandemic virus. Science 310, 77–80 (2005).
Crossref
PubMed
Google Scholar
40
YN Lee, PD Bieniasz, Reconstitution of an infectious human endogenous retrovirus. PLoS Pathog 3, e10 (2007).
Crossref
PubMed
Google Scholar
41
DG Gibson, et al., Complete chemical synthesis, assembly, and cloning of a Mycoplasma genitalium genome. Science 319, 1215–1220 (2008).
Crossref
PubMed
Google Scholar
42
WC McRoy, RS Baric, Amino acid substitutions in the S2 subunit of mouse hepatitis virus variant V51 encode determinants of host range expansion. J Virol 82, 1414–1424 (2008).
Crossref
PubMed
Google Scholar
43
RA Lamb, TS Jardetzky, Structural basis of viral invasion: Lessons from paramyxovirus F. Curr Opin Struct Biol 17, 427–436 (2007).
Crossref
PubMed
Google Scholar
44
JH Schickli, LB Thackray, SG Sawicki, KV Holmes, The N-terminal region of the murine coronavirus spike glycoprotein is associated with the extended host range of viruses from persistently infected murine cells. J Virol 78, 9073–9083 (2004).
Crossref
PubMed
Google Scholar
45
YY Mitsuki, et al., A single amino acid substitution in the S1 and S2 Spike protein domains determines the neutralization escape phenotype of SARS-CoV. Microbes Infect 10, 908–915 (2008).
Crossref
PubMed
Google Scholar
46
MA Hofmann, SD Senanayake, DA Brian, A translation-attenuating intraleader open reading frame is selected on coronavirus mRNAs during persistent infection. Proc Natl Acad Sci USA 90, 11733–11737 (1993).
Crossref
PubMed
Google Scholar
47
W Ren, et al., Difference in receptor usage between severe acute respiratory syndrome (SARS) coronavirus and SARS-like coronavirus of bat origin. J Virol 82, 1899–1907 (2008).
Crossref
PubMed
Google Scholar
48
W Li, et al., Efficient replication of severe acute respiratory syndrome coronavirus in mouse cells is limited by murine angiotensin-converting enzyme 2. J Virol 78, 11429–11433 (2004).
Crossref
PubMed
Google Scholar
49
SA Kopecky-Bromberg, et al., Severe acute respiratory syndrome coronavirus open reading frame (ORF) 3b, ORF 6, and nucleocapsid proteins function as interferon antagonists. J Virol 81, 548–557 (2007).
Crossref
PubMed
Google Scholar
50
SA Kopecky-Bromberg, L Martinez-Sobrido, P Palese, 7a protein of severe acute respiratory syndrome coronavirus inhibits cellular protein synthesis and activates p38 mitogen-activated protein kinase. J Virol 80, 785–793 (2006).
Crossref
PubMed
Google Scholar
51
M Frieman, et al., Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. J Virol 81, 9812–9824 (2007).
Crossref
PubMed
Google Scholar
52
MG Wathelet, M Orr, MB Frieman, RS Baric, Severe acute respiratory syndrome coronavirus evades antiviral signaling: Role of nsp1 and rational design of an attenuated strain. J Virol 81, 11620–11633 (2007).
Crossref
PubMed
Google Scholar
53
RL Graham, et al., The nsp2 replicase proteins of murine hepatitis virus and severe acute respiratory syndrome coronavirus are dispensable for viral replication. J Virol 79, 13399–13411 (2005).
Crossref
PubMed
Google Scholar
VIEW FULL TEXT|DOWNLOAD PDF
Further reading in this issue
RESEARCH ARTICLEDECEMBER 16, 2008
Conformal geometry of the retinal nerve fiber layer
P. Juhani Airaksinen, Stephen Doro, Jukka Veijola,
RESEARCH ARTICLEDECEMBER 16, 2008
Dynamical roles of metal ions and the disulfide bond in Cu, Zn superoxide dismutase folding and aggregation
Feng Ding, Nikolay V. Dokholyan and
RESEARCH ARTICLEDECEMBER 16, 2008
Alphavirus-derived small RNAs modulate pathogenesis in disease vector mosquitoes
Kevin M. Myles, Michael R. Wiley, [...]Zach N. Adelman and
Trending
RESEARCH ARTICLEJANUARY 8, 2024
Hormonal basis of sex differences in anesthetic sensitivity
Half of all surgical patients receiving anesthesia are females. Anesthetics affect sexually dimorphic brain regions involved in sleep and arousal. Yet, sex differences and the effects of sex hormones on anesthetic sensitivity have not been ...General anesthesia—a pharmacologically induced reversible state of unconsciousness—enables millions of life-saving procedures. Anesthetics induce unconsciousness in part by impinging upon sexually dimorphic and hormonally sensitive hypothalamic circuits ...
Andrzej Z. Wasilczuk, Cole Rinehart, [...]ReCCognition Study Group,
[ This page built with JPICedar v. j022924a Fri, 4-12-2024 Time : 4:30p]