BC SCIENCE
Saved by @CodyyyGardner

Let Uncle Ralph educate you
Synthetic Viral Genomics:
Risks and Benefits for Science and Society

 "Alternatively, “No See’m” sites can be used to insert foreign genes into viral, eukaryotic, or microbial genome or vector, simultaneously removing all evidence of the restriction sites that were used in the recombinant DNA manipulation"
By orientating the restriction sites as “No See’m”, the sites are removed during reassembly, leaving only the desired mutation in the final DNA product.
The dual properties of strand specificity & a variable end overhang that can be tailored to match any sequence allow for Esp3I sites to be engineered as “universal connectors” that can be joined with any other four nucleotide restriction site overhangs (e.g. EcoRI, PstX1, BamH1)
Seamless assembly (also called No See’m Sites (85)) cascades have been used to assemble full length cDNAs of the coronaviruses mouse hepatitis virus, transmissible gastroenteritis virus, infectious bronchitis virus and SARS-CoV (Refs: 85,86,87)
Type IIS restriction endonucleases recognize asymmetric binding sites & leave asymmetric ends
These enzymes can be used to create unique interconnecting junctions, which can be subsequently removed from final assembly product allowing seamless reconstruction of an exact sequence
With enzymes like Esp3I, interconnecting restriction site junctions can be located at ends of each cDNA & systematically removed during assembly of complete full-length cDNA product
Consequently, knowledgeable experts can theoretically reconstruct full length synthetic genomes for any of the high priority virus pathogens, although technical concerns may limit the robustness of these approaches.
Another approach might be to “humanize” zoonotic viruses by inserting mutations into virus attachment proteins or constructing chimeric proteins that regulate virus species specificity (viral attachment proteins bind receptors, mediating virus docking and entry into cells).
The pathogenicity of these chimeric coronaviruses is unknown
The pathogenicity of these chimeric coronaviruses is unknown
The pathogenicity of these chimeric coronaviruses is unknown
The pathogenicity of these chimeric coronaviruses is unknown
"Scapegoat option"
This powerful technique provides bioterrorists with a “scapegoat” option; leaving a sequence signature that misdirects efforts at tracking the true originators of the crime. Even better, the approach could be used to build mistrust &/or precipitate open warfare
Dual Use
Uncle Ralph summarises his findings (1)
Uncle Ralph Concludes (2)
Hence Ecohealth DARPA/DTRA spooks & virus thieves collaboration with Baric (UNC) Lipkin (Mailman) NIchols (Atlanta CDC) and USAMRIID (Bavari, Totura et al) & Jonathan Epstein's palpable concern about dual use references in the @USRightToKnow FOIA emails
A scary read for an experimental monkey!
Delving into the cold and calculating mind of a twisted genius?
https://t.co/MrwxYK93cx
Bring Uncle Ralph and his transgenic mice in for questioning!
unroll @threadreaderapp
You just read the Baric Paper, one of 6 in a series, the rest are available here in a 191 page document
https://t.co/rjNZ8aDNDl
Related Papers
1. Synthetic Genomics: Options for Governance (2008) https://t.co/A32jEfL1TQ
2. Sequence Screening - Robert Jones (2005)
https://t.co/zOqMroYbVU
3. Synthetic Biology as a Field of Dual-Use Bioethical Concern - Alexander Kelle
https://t.co/Va1movPqvs
Related papers (2)
4. Sanghvi Y. A Roadmap to the Assembly of Synthetic DNA from Raw Materials.
https://t.co/4kWu4vzcXC
5. Collett MS. Impact of Synthetic Genomics on the Threat of Bioterrorism with Viral Agents.
https://t.co/OcaTmuCUBy
Related Papers (3)
6. Fleming DO. Risk Assessment of Synthetic Genomics: A Biosafety & Biosecurity Perspective.
https://t.co/JBcLVLr005
7. Risk Governance of Synthetic Biology
https://t.co/9BE9X5s0Fb
8. US Competitiveness in Synthetic Biology
https://t.co/OCZJdsXSjl
Related papers (4)
9. Ensuring security of synthetic biology
https://t.co/7kPE1OpbrS
10. Synthetic biology: emerging research field in China
https://t.co/T3BcaDvUsa
11.
What rough beast? Synthetic biology, uncertainty,& the future of biosecurity (2016)
https://t.co/XTlBXILEWh
Back to Baric et Al
Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus (2003)
https://t.co/k8PI1rbcm8
Systematic Assembly of a Full-Length Infectious Clone of Human Coronavirus NL63 (2008)
https://t.co/bWhAv1wsC0
For the Record (WIV)
In 2016, Shi and her team at the WIV, in conjunction with the New York-based EcoHealth Alliance, constructed a full-length clone of a bat coronavirus called SL-CoV WIV1. They assembled it in discrete segments.
They genetically engineered the virus using the pGEM®-T Easy Vector Systems to join the segments. This system, also available on the internet, gives researchers several options for how to remove GM inserts that can be seen as signatures of a lab-made virus.18
pGEM®-T Easy Vector Systems:
“Thus, several options exist to remove the desired insert DNA with a single restriction digestion.”
This shows that researchers at the WIV have the ability to genetically engineer viruses and remove the signatures of the genetic engineering.
WIV & EcoHealth Alliance published a paper in 2017 on how they genetically modified spike proteins of 8 bat coronaviruses, by cutting & pasting genetic material from other coronaviruses, so that the viruses infected the human ACE2 receptor
They used pGEM®-T Easy Vector Systems to join the segments to genetically engineer these viruses.
They showed how they can insert new spikes into viruses. The researchers state:
“Then any spike could be substituted into the genome of SARSr-CoV WIV1 through this strategy.”
This shows that researchers at WIV have the ability to genetically modify multiple coronaviruses to insert new spikes, and these new viruses cannot be detected as genetically engineered.
The research clearly shows that GOF researchers at WIV could assemble SARS-CoV-2 from bat coronaviruses, such as RaTG13 or similar & spike protein from Malayan Pangolins & insert multi-basic cleavage sites into precise regions of spike & leave no evidence of genetic engineering.
All the above about WIV research is taken verbatim from an excellent article by Andre Leu on the website of @OrganicConsumer
COVID 19: The Spike and the Furin Cleavage
https://t.co/bDOMmhpJ1t

More from Billy Bostickson 🏴👁&👁 🆓

Billy Bostickson \U0001f3f4\U0001f441&\U0001f441 \U0001f193
Billy Bostickson 🏴👁&👁 ...
@BillyBostickson
1/X Bellagio Rockefeller Initiative (Global Virome Project)
Another look at some key people in the GVP & their media statements since January 2020 about "natural origin" and impossibility of a "lab leak"
Full List of 18 scientists in photograph is here:
https://t.co/yHoAjwQjI8


2/X
We have already looked closely at Dennis Carroll, Peter Daszak, George Gao, W. Ian Lipkin & seen their public statements supporting "natural origin" & hysterical attacks on "lab leak" theories.
This thread focuses on:
Michael Kurilla
Jonna Mazet
Edward Rubin (Metabiota)

3/X
If we look at the leadership team of GVP, you will see some familiar faces:
https://t.co/WdT8sNnfmy
Most familiar is probably Peter Daszak, the wealthy Treasurer, also with the key role of Secretary.
Note: USAID PREDICT Project - EcoHealthAlliance


4/X
The "Chair" is Dr. Dennis Carroll
We will have a look at his media statements on the coronavirus pandemic by inputting his name in Twitter search box (very easy!)
Note: Previously was Director of USAID Emerging Pandemic Threats Program


5/X Dennis Carroll's video media statements (1):
CNN
(note Qinghai Thangka positioned in background which Daszak also shows off in his videos obtained during EcoHealth virus hunt with
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Billy Bostickson \U0001f3f4\U0001f441&\U0001f441 \U0001f193
Billy Bostickson 🏴👁&👁 ...
@BillyBostickson
1. WIV Research groups (archived pages) for Reference
dsRNA viruses molecular biology

https://t.co/lwrQoo6ygG


2. Emerging Viruses Group

https://t.co/ND56gVGOAn


3. Structural Biology of Viral Genome Replication
https://t.co/wUqc0YIP3X


4. Molecular Biology of hepatitis Viruses & Gene Therapy
https://t.co/snBbxkhNsC


5. Insect Virus Genetic Engineering Lab
https://t.co/S68mVOA2Ob
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Billy Bostickson \U0001f3f4\U0001f441&\U0001f441 \U0001f193
Billy Bostickson 🏴👁&👁 ...
@BillyBostickson
PCR Drosten Fail Compilation
1. Ten Fatal Errors: Scientists Attack Paper That Established Global PCR Driven Lockdown
https://t.co/25p88F2xu6
2. Review report Corman-Drosten et al.
https://t.co/8rlT4MWoyI
3. Consequences of false

PCR Drosten Fail Compilation
4. Files for download
https://t.co/SqDrZjB9em
5. Addendum
https://t.co/IygCWvnAsP
6. False positives in reverse transcription PCR testing for SARS-CoV-2
https://t.co/HxpSZkzZ7Q
7. Discussion on Retraction

PCR Drosten Fail Compilation
8. 124 Tweet Thread by Bobby Rajesh Malhotra
@Bobby_Network
https://t.co/cdot5yIeuf
9. Another shorter thread by @Bobby_Network
https://t.co/kqFW9GsSeD
10. Thread by Abir Ballan

PCR Drosten Fail Compilation
11. Mini Thread by @Kevin_McKernan
https://t.co/pntydnd80L
12. Another longer thread by @Kevin_McKernan
https://t.co/5hBYIlXFnQ
13.
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Billy Bostickson \U0001f3f4\U0001f441&\U0001f441 \U0001f193
Billy Bostickson 🏴👁&👁 ...
@BillyBostickson
1. Thread: RaTG13 Real or Fake?
A Norwegian Scientist @Vehuardo recently claimed RaTG13 is an "authentic virus"
https://t.co/tAXMNTwdhA
This failed to take into account the work of our DRASTIC colleagues & other published analyses.
This thread is a polite attempt to educate.

2. https://t.co/tw606vgHwp
which says RaTG13 doesn't make sense on how it was sequenced, or how the data got so messed up in the lab as to include copy paste mistakes, bat dna, pangolin dna, rice dna, mice dna, & nobody should use it to form conclusions until it's properly vetted

2 (cont)
"sequence is incomplete & provided segment doesn't function. The identified markers suggest partial match basis & actual virus itself as described by the sequence can't functionally exist due to incoherent structures that appear to be the result of sample contamination"


3. RaTG13 integrity is further attacked here:
The Validity of critical pieces of evidence for the natural origin of SARS-CoV-2 is Dubious, and needed to be

4. and here:
Major Concerns on the Identification of Bat Coronavirus Strain RaTG13 and Quality of Related Nature
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Billy Bostickson \U0001f3f4\U0001f441&\U0001f441 \U0001f193
Billy Bostickson 🏴👁&👁 ...
@BillyBostickson
Meat is high in Glycosaminoglycans
(central to coronavirus's ability to attach to host cells)
Beef, pork poultry, seafood are rich in heparan sulfate (glycosaminoglycans), highly charged anchors for SARSCoV2
Meat surfaces critical factors for transmission
https://t.co/yP6lH22OTF

1/5 Science behind that tweet just in case anyone thinks I speak without awareness ;)
Glycosaminoglycans from fresh water fish processing discard - Isolation, structural characterization, and osteogenic activity
https://t.co/fyjac03IuV
SALMON MOUTHS

2/5 Salmon Mouths 2
Characterization of Glycosaminoglycans in Gaping and Intact Connective Tissues of Farmed Atlantic Salmon (Salmo salar) Fillets by Mass

3/5 Why does SARS-COV-2 like meat & seafood?
Glycosaminoglycan binding motif at S1/S2 proteolytic cleavage site on spike glycoprotein may facilitate novel coronavirus (SARS-CoV-2) host cell entry
https://t.co/DpHF1WmxeG
Glycosaminoglycans and

4/5 Glycosaminoglycans
Role of in Infectious Disease
(same authors as above)
https://t.co/3q5dn2Io3Q
Glycosaminoglycans in Infectious Disease
https://t.co/Dep7aJlpLP
Murine Coronavirus with an Extended Host Range Uses Heparan Sulfate as an Entry
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DNA consists of four different 'bases', A, C, G and T. These bases have specific meaning within our biology. Specifically, within the 'coding part' of a gene, a triplet of bases encodes for an amino acid


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https://t.co/jD4cMt0PJ0


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