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 🏴👁&👁 🆓

@snip96581187 @Daoyu15 @lab_leak @walkaboutrick @ydeigin @Ayjchan @franciscodeasis @TheSeeker268 @angie_rasmussen Clearly, because as I have been saying for 8 months now, DTRA and DARPA have been using Ecohealth and UC Davis to collect novel pathogens for gain of function work back in the USA. I have documented this in many threads which I will post here just to annoy everyone.

@Daoyu15 @lab_leak @walkaboutrick @ydeigin @Ayjchan @franciscodeasis @TheSeeker268 @angie_rasmussen


@Daoyu15 @lab_leak @walkaboutrick @ydeigin @Ayjchan @franciscodeasis @TheSeeker268 @angie_rasmussen


@Daoyu15 @lab_leak @walkaboutrick @ydeigin @Ayjchan @franciscodeasis @TheSeeker268 @angie_rasmussen


@Daoyu15 @lab_leak @walkaboutrick @ydeigin @Ayjchan @franciscodeasis @TheSeeker268 @angie_rasmussen
1. Project 1742 (EcoHealth/DTRA)
Risks of bat-borne zoonotic diseases in Western Asia

Duration: 24/10/2018-23 /10/2019

Funding: $71,500
@dgaytandzhieva
https://t.co/680CdD8uug


2. Bat Virus Database
Access to the database is limited only to those scientists participating in our ‘Bats and Coronaviruses’ project
Our intention is to eventually open up this database to the larger scientific community
https://t.co/mPn7b9HM48


3. EcoHealth Alliance & DTRA Asking for Trouble
One Health research project focused on characterizing bat diversity, bat coronavirus diversity and the risk of bat-borne zoonotic disease emergence in the region.
https://t.co/u6aUeWBGEN


4. Phelps, Olival, Epstein, Karesh - EcoHealth/DTRA


5, Methods and Expected Outcomes
(Unexpected Outcome = New Coronavirus Pandemic)
"The coronavirus in Italy since September 2019"
Unexpected detection of SARS-CoV-2 antibodies in the prepandemic period in Italy
First Published November 11, 2020
https://t.co/nhlccks5AS
Smashed Paywall thanks to @Sci_Hub
https://t.co/wWPFJMjjFJ

Maybe a coincidence but an agreement was recently signed by the authors' institute and guess who?
https://t.co/vh7qX3hyPD


The Lead Author specifically of the paper:
"Unexpected detection of SARS-CoV-2 antibodies in the prepandemic period in Italy"
is Giovanni Apolone of the "Fondazione IRCCS Istituto Nazionale Tumori" Milan, Italy


Some funding is involved....
Anyone good at Italian feel free to comment?


"You findee sars cov 2 in old sample back September time, we give you big fat red envelopeee, okey dokey?"

More has emerged regarding the Chinese connection
"Oncology research, Italy and China close to the collaboration protocol"
.https://t.co/Mt5hO9Dggc

More from Science

@mugecevik is an excellent scientist and a responsible professional. She likely read the paper more carefully than most. She grasped some of its strengths and weaknesses that are not apparent from a cursory glance. Below, I will mention a few points some may have missed.
1/


The paper does NOT evaluate the effect of school closures. Instead it conflates all ‘educational settings' into a single category, which includes universities.
2/

The paper primarily evaluates data from March and April 2020. The article is not particularly clear about this limitation, but the information can be found in the hefty supplementary material.
3/


The authors applied four different regression methods (some fancier than others) to the same data. The outcomes of the different regression models are correlated (enough to reach statistical significance), but they vary a lot. (heat map on the right below).
4/


The effect of individual interventions is extremely difficult to disentangle as the authors stress themselves. There is a very large number of interventions considered and the model was run on 49 countries and 26 US States (and not >200 countries).
5/
This is a thread on statistics in science: 1/7 (via @LogicofScience)

Basic Statistics Part 1: The Law of Large Numbers https://t.co/wUH8eAAIak

#Science #Statistics


Basic Statistics Part 2: Correlation vs. Causation

https://t.co/Azhyl8pDsX (2/7)

Basic Statistics Part 3: The Dangers of Large Data Sets: A Tale of P values, Error Rates, and Bonferroni Corrections

https://t.co/LetN6aEBRM (3/7)

Basic statistics part 4: understanding P values

https://t.co/K8MMMgTCOf (4/7)

Basic Statistics Part 5: Means vs Medians, Is the “Average”

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