mod-mRNA "Vaccines", DNA Fragment Risks

Don't give me no lies and keep your strands to yourself

Subtitle is a shoutout to the Southern rock group the Georgia Satellites and their one hit wonder “Keep Your Hands to Yourself"

“Absence of proof is not proof of absence”

“I’m from Missouri, the show me state. Show me the data”

latter quote from Dr. Phil Needleman, author of the 10 Commandments of Drug Development

Executive Summary.

After three years of drug manufacturers and regulatory agencies insisting the COVID shots were safe, study findings by independent scientists now demonstrate that certain modified-mRNA shots may meet or exceed regulatory standards for “adulteration” with DNA fragments.  In the case of the Pfizer/BioNTech product, these fragments appear to include DNA of a particular sequence (SV40 origin-enhancer-promoter) which is biologically active in animal cells.  The presence of SV40-derived DNA fragments was not fully disclosed by the manufacturers to regulatory authorities. 

This lack of transparency raises serious questions about how these fragments and specific DNA sequences escaped regulatory consideration, and why the presence and risks of significant amounts of SV40- and bacterial-derived small DNA fragments were not discussed by the manufacturers in documents submitted to regulators. The study’s findings have been replicated by other scientists prompting several international medical organizations to call for the immediate recall of all COVID shots. 

To be clear, these studies have not found either full-length SV40 DNA, SV40 virus, or the cancer-associated SV40 “Large T antigen” sequences among the contaminants. 

FDA, the European Medicines Agency (EMA) and Health Canada assert that there are no risks associated with this DNA fragment adulteration, willfully overlooking the decades-long rich body of peer reviewed literature documenting that both short RNA fragments and DNA fragments (oligonucleotides) can be used to regulate gene expression, and DNA fragments can insert and recombine with genomic DNA in cells, embryos, and tissues. In a wide range of experimental models, insertion and recombination of foreign DNA is associated with both cancer and birth defects.

FDA, EMA and Health Canada also overlook that the self-assembling lipid nanoparticle technology employed to deliver spike-coding mod-mRNA is indiscriminate, and will self-assemble around mod-mRNA, short RNA fragments, and short DNA fragments, efficiently delivering all of these molecules to the cells and tissues of those patients who accept (or are enticed, compelled or coerced to accept) these products. Transfected tissues throughout the body will include ovarian tissue in female patients of all ages, based on the Pfizer non-clinical study data disclosed in their initial common technical document submission.

Background.

Many have now seen the data of Speicher et al, currently summarized in a pre-print article, which demonstrates the adulteration (or undisclosed contamination) of injected Pfizer/BioNTech and Moderna COVID-19 pseudo-mRNA vaccine products with short DNA fragments. In both of these products there are previously undisclosed substantial amounts of short linear DNA fragments derived from degrading the DNA templates used for manufacturing the pseudo-mRNA biological component of the complex self-assembling lipid nanoparticles. See “What is Adulteration of pseudo-mRNA vaccines, and why should you care?” for further details.

Just to review, in the case of the Pfizer/BioNTech product, these DNA fragments include sequences derived from the origin of replication (“ori”) and promoter/enhancer sequences (strong genetic switch elements) found in the circular DNA genome of the SV40 virus. Simian Virus 40 (SV40) is causally associated with solid tumors (sarcomas) and did contaminate early live attenuated (Sabin) polio vaccines, but full SV40 is not contaminating the mod-mRNA vaccines.

To round out the SV40 story, the majority of the cancer-associated activity in SV40 has been localized to the “large T antigen” protein, and there are no large T antigen sequences included in any of the Pfizer/BioNTech DNA fragments. Which is not to say that there is no cancer risk - more about that below.

Bona fides.

From time to time I am reprimanded by social media commentators and trolls to “stay in your lane”. I generally disregard these comments because my “lane” has broadened over the last four years, and our scope of knowledge and competency has increased as Jill and I have transitioned to becoming full time policy analysts and writers. But in the case of issues relating to DNA and RNA delivery, genomic insertional mutation risks, activation/inactivation of oncogenes, tumor suppressor genes and general mutagenesis, this has long been in my expertise portfolio. In fact, long ago Jill and I helped launch a breast cancer research program for the US Army and Uniformed Services University of the Health Sciences!

Beginning in 1983, I began my journey in learning the trade and craft of molecular virology in a UC Davis School of Medicine Pathology lab which focused on the molecular biology of breast cancer and the use of the Mouse Mammary Tumor Virus retrovirus to determine linkages between oncogenes, tumor suppressor genes, retroviral insertions (ergo “integration”), and murine breast cancer. This was under the direction of Pathology Professor Dr. Robert Cardiff, MD, PhD, one of the early molecular pathologists, who had just completed a sabbatical at UCSF in the laboratories of Drs. Bishop and Varmus (Nobel laureates for discovery of oncogenes). This was where I learned to extract, purify, handle and analyze both DNA and RNA from tumor and blood samples.

The way that many oncogenes and tumor suppressor genes (and many other key regulatory proteins) were first identified was to introduce genetic elements (retroviruses, transposons - “jumping genes”, or other DNA sequences) into the cells or tissues of animal models. Another way was to take naturally occurring cancers (such as chicken sarcomas), grind them up and look for tumor-associated viruses that might have picked up genetic material from the host. From this you can appreciate that pretty much anything that interferes with the integrity of animal genome DNA by inserting itself into chromosomal DNA can cause cancer.

One of the hot issues relating to the manufacturing process and purity/adulteration of the mod-mRNA vaccines is the presence and levels of endotoxin, a cell wall molecule derived from the bacteria used to manufacture the DNA. Humans are quite sensitive to endotoxin, and pharmaceutical contamination of injectable products with endotoxin is strongly associated with anaphylactic shock, a known adverse event associated with the current mod-mRNA products. I believe that I was the first to take this issue head on for DNA/lipid nanoparticle complex manufacturing (Improved method for the removal of endotoxin from DNA, P M Montbriand , R W Malone. J Biotechnology. 1996 Jan 26;44(1-3):43-6). Also relevant to the current cationic lipid nanoparticle formulation manufacturing is incorporation of cholesterol to improve the fluidity, another advance in the art pioneered in my laboratory (Cholesterol enhances cationic liposome-mediated DNA transfection of human respiratory epithelial cells. Bennett MJ, Nantz MH, Balasubramaniam RP, Gruenert DC, Malone RW. Bioscience Reports 1995 Feb;15(1):47-53.)

So there is that. Bona fides established. These issues are most definitely in my lane. Now lets turn back to contamination of pseudo-mRNA vaccines with short DNA fragments (oligonucleotides). This will get a bit technical, because it is necessary to counter FDA, EMA and Health Canada propaganda with facts and data.

European Medicines Agency (EMA), FDA and Health Canada Correspondence

Seeking clarification of official regulatory positions regarding the newly identified DNA fragment contamination/adulteration, the journalists Mary Beth Pfeiffer and Zachary Stieber have submitted many questions and requests for information to FDA, the European Medicines Agency (EMA) and Health Canada regulatory communications offices. Both reporters have indicated to me that the regulatory agencies refused to address many of their questions.

Mary Beth Pfeiffer’s recent article covering the Health Canada response can be found here at Trial Site News.

DNA Fragment Won’t Cause Cancer, Health Canada Says. Says Substance Is 'Inactive, Not Functional.' By Mary Beth Pfeiffer Oct. 27, 2023

I will hold commentary until after the regulatory response to allow the reader to review and interpret the results of each interaction by him or herself. Italics are used to indicate reporter questions in the lengthy EMA correspondence.

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European Medicines Agency (EMA)

From: Zack Stieber
Sent: 27 October 2023 08:39
To: Press <Press@ema.europa.eu>
Subject: (HC) Request for Comment

Hello,

Good morning.

Has EMA confirmed the presence of a SV40 sequence in Pfizer's COVID-19 vaccine?

An EMA spokesperson said earlier this year that "there is no evidence to indicate the presence of SV40...in the formulation of COVID-19 vaccines."

How did EMA learn of this sequence's presence, and when did it learn of it? Did Pfizer ever disclose the sequence to EMA?

An SV40 sequence is present in the DNA plasmid starting material of Comirnaty. In this case the sequence is not directly relevant for plasmid production in E. coli or for the mRNA production process so it is considered to be a non-functional part of the structure of the source plasmid.

SV40 is a naturally occurring virus. The virus itself is not used in the manufacture of the vaccine. Specific sequences for the non-infectious parts of SV40 are commonly present in plasmids used for manufacturing of biological active substances. The sequence for non-infectious parts of SV40 is only a small fraction of the entire SV40 sequence.

During the manufacturing process, this sequence and other plasmid DNA sequences are broken down and removed. Fragments of the SV40 sequence may only be present as residual impurities at very low levels that are routinely controlled. There is no scientific evidence that any of these SV40 fragments can act as insertional mutagens. While the full DNA sequence of the plasmid starting material was provided in the initial marketing authorisation application for Comirnaty, the applicant did not specifically highlight the SV40 sequence, as it was considered to be a non-functional part of the plasmid. They have since clarified this information in response to questions raised by EMA.

Finally, is EMA concerned about the possible safety concerns over the sequence's inclusion?

EMA has set appropriate limits for the level of residual DNA in the vaccines.

The manufacturing process of mRNA vaccines is carefully designed and controlled to ensure that the level of residual DNA is below acceptable and safe levels.

We can confirm that we have not seen any reliable evidence of residual DNA exceeding approved/safe levels for either of the EU-authorised mRNA vaccines, Comirnaty or Spikevax.

EMA has seen no evidence of an association between mRNA vaccines and adverse events that could be linked to the presence of DNA material, nor are we aware of any scientific evidence showing that the very small amounts of residual DNA that may be present in vaccine batches could integrate into the DNA of vaccinated individuals.

EMA and EU Member States continuously monitor the safety of all COVID-19 vaccines to ensure any possible risks are detected and managed as early as possible, and EMA carefully assesses suspected side effects to determine if they could have been caused by a vaccine. We have recently published a report on medicine safety monitoring during the pandemic (https://www.ema.europa.eu/en/news/report-how-eu-ensured-safety-medicines-during-covid-19).

Thank you,

Zack Stieber
The Epoch Times

Commentary:

Is this statement accurate - “The manufacturing process of mRNA vaccines is carefully designed and controlled to ensure that the level of residual DNA is below acceptable and safe levels.”? Currently available information indicates that, as a matter of established fact and public record, the current manufacturing process (Process 2) was hastily developed and implemented when the process employed to produce the initial clinical trial material was not able to support the manufacturing requirements. This statement by EMA is clearly disingenuous at best; basically just propaganda.

The first two paragraphs of response regarding the SV40 Origin of Replication-Enhancer-Promoter sequences included in the Pfizer/BioNTech product Comirnaty are not technically correct and include an internal inconsistency. The statement “the sequence is not directly relevant for plasmid production in E. coli” appears to be incorrect. The sequence in question drives production of the RNA coding for the protein which confers bacterial resistance to Kanamycin/Neomycin, which is used to maintain the plasmid in the bacteria which manufacture it during their growth. It is also used to maintain this type of “shuttle vector” plasmid in animal cells for experimental purposes. These sequences are therefore biologically active in both bacterial and animal cells, and are directly relevant to plasmid production. This begs the question of why these sequences are even present in this plasmid, as a well designed plasmid would have removed these sequences to minimize plasmid size and maximize plasmid yield (and stability), and would have exclusively employed a more acceptable antibiotic resistance cassette. The statement “Specific sequences for the non-infectious parts of SV40 are commonly present in plasmids used for manufacturing of biological active substances.” also reveals the contradictory fallacy in the response. If commonly used, then why are they included if they serve no function?

One interpretation of this anomaly involving presence of SV40 origin of replication and strong regulatory sequences is that this plasmid was hastily selected rather than bespoke for this manufacturing process (process 2) because the PCR-based process 1 could not support the required manufacturing throughput. So this plasmid may have been basically pulled off of a research project for use by the manufacturing process development team. That is the most benign explanation I can come up with.

The statement “During the manufacturing process, this sequence and other plasmid DNA sequences are broken down and removed” is at the heart of the adulteration concerns. The manufacturing process asserted to guarantee that these “plasmid DNA sequences are broken down and removed” is not actually removing the plasmid DNA sequences. If it was, then we would not be having this discussion. It may be reducing the contamination, but it is not removing it. A more transparent and accurate answer would be “plasmid DNA sequences are broken down and removed to what EMA considers to be a safe and acceptable level”. Sloppy at best. One is left wondering if the grown ups are minding the EMA (or Paul Erlich Institute) regulatory shop.

This next statement is really the absolute crux of the whole matter. We have established from this correspondence that EMA is aware that there are, in fact, contaminating DNA fragments, but that the EMA asserts that the level of contamination is safe. Then this rather odd assertion is provided: “There is no scientific evidence that any of these SV40 fragments can act as insertional mutagens.” The absence of evidence does not provide evidence of absence. This statement inverts standard regulatory policies and procedures, and is akin to a regulatory authority allowing the manufacturer to play “catch me if you can”. The proper regulatory position when the normal tension between regulatory authority and manufacturer/sponsor is maintained would have yielded the statement “rigorous long term genotoxicity and insertional mutagenesis studies have been completed and have demonstrated an acceptably low minimal risk of genomic modification.”

Then we have the statement that “EMA has set appropriate limits for the level of residual DNA in the vaccines.” Not provided are either the limits which have been set, nor any information on how they were set or what the data to support those limits are. At this point, given the history of rushed, sloppy regulatory practices relating to the mod-mRNA vaccine approval and pharmacovigilance processes, the default assumption (unless proven otherwise) must be that these levels were established in an arbitrary and capricious manner without rigorous non-clinical safety testing to technically define the acceptable level of DNA fragment contamination.

Here is the specific guidance from the FDA concerning genotoxicity studies (see the link for the PDF of the technical guidance document):

S2(R1) Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use

JUNE 2012

This guidance combines and replaces two ICH guidances, S2A Specific Aspects for Regulatory Genotoxicity Tests for Pharmaceuticals (ICH S2A guidance) and S2B Genotoxicity: A Standard Battery for Genotoxicity Testing of Pharmaceuticals (ICH S2B guidance). The purpose of the revision is to provide guidance on optimizing the standard genetic toxicology battery for prediction of potential human risks, and on interpreting results, with the goal of improving risk characterization for carcinogenic effects that have their basis in changes in the genetic material. The revised guidance describes internationally agreed-upon standards for follow-up testing and interpretation of positive results in vitro and in vivo in the standard genetic toxicology battery, including assessment of nonrelevant findings. This guidance is intended to apply only to products being developed as human pharmaceuticals.

There are no specific safety, genotoxicity or insertional mutagenesis guidances developed by EMA, FDA or Health Canada for mod-mRNA vaccines. This is highly unusual, and reflects the rush to approve these products.

In contrast, there has been extensive and rigorous study of insertional mutagenesis risks associated with the vast majority of other gene therapy technologies. There has also been extensive and rigorous study and guidance developed for plasmid DNA vaccines. The history of that guidance has been summarized in a 2010 peer reviewed article titled “FDA Guidance on Prophylactic DNA Vaccines: Analysis and Recommendations”. This particular DNA vaccine guidance appears to me to be the closest thing currently available covering the proper evaluation of risks which might be associated with a new non-viral delivery formulation technology and its use to deliver both mod-mRNA as well as contaminating DNA fragments. The following summary is particularly relevant:

3.4) Plasmid Integration issues

DNA is considered a contaminant of “conventional” (non-DNA) vaccines. Manufacturers typically minimize or remove extraneous DNA during the production process to minimize human exposure to potentially injurious material. Not surprisingly, concerns that plasmid DNA might integrate into the host genome, increasing the likelihood of malignant transformation, genomic instability, or cell growth dysregulation were raised when DNA vaccines were first proposed for clinical use [31;36;41]. Based on advice from the Vaccine's Advisory Committee, the FDA set guidelines designed to insure that the frequency of plasmid integration would be substantially lower than the spontaneous mutation rate [42].

Initial efforts to examine whether DNA vaccines could integrate into the host genome involved monitoring the biodistribution and persistence of plasmids in vivo. In most studies, DNA was isolated from tissues of vaccinated animals and the presence of plasmid examined using sensitive Q-PCR assays. Integration was then assessed by size fractionating high molecular weight genomic DNA free from smaller unintegrated plasmids (this sometimes required restriction endonuclease digestion to eliminate plasmid concatemers) [24;43;44]. Q-PCR and/or RAIC-PCR were then used to detect and quantify residual plasmid present in the genomic DNA preparation. Results showed that conventional intra-muscular, subcutaneous, intra-dermal, and particle-mediated delivery of DNA plasmids rarely resulted in the long-term persistence of plasmid in tissues distal from the site of vaccine administration [24;43-46]. In contrast, tissue at or near the site of administration (such as muscle and the overlying skin) commonly contained thousands or even millions of copies of plasmid per microgram of host DNA shortly after delivery [24;43-49]. Over time, the vast majority of this plasmid disappeared [43-49]. Based on studies involving multiple different plasmid backbones, and of the same plasmid backbone with various inserts, the revised FDA Guidance document indicated that biodistribution/ persistence studies could be waived for vaccines prepared using a plasmid vector previously documented to have an acceptable biodistribution/integration profile. For novel plasmids or methods of formulation and delivery, integration studies would be required when plasmid persists at levels exceeding 10,000 copies per ug of host DNA. This recommendation reflects the expectation that only a tiny fraction of persisting plasmid will integrate into the host genome, regardless of the method of delivery [45].

In evaluating the potential harm of plasmid integration, it should be noted that the risk of introducing plasmids with strong regulatory regions into the host genome far exceeds that associated with random point mutations [43;50]. Moreover, the technology used to detect plasmid persistence does not examine the frequency with which short fragments of plasmid integrate. In this context, sections of DNA as short as 7 bp can affect rates of integration or recombination. Examples include the VDJ recombination signal sequence and related sequences, chi-like elements and minisatellites, ALU sequences, a recombinase signal present in hepatitis B and mammalian genomes, and topoisomerase II recognition sites [43].

Each of the cited references refer to the numbered citation in the source article.

The following references are particularly relevant:

Ledwith BJ, Manam S, Troilo PJ, Barnum AB, Pauley CJ, Nichols WW. Plasmid DNA vaccines: Assay for integration into host genomic DNA. Dev Biol. 2000;104:33–43

Smith HA. Regulation and Review of DNA Vaccine Products. Dev Biol. 2000;104:57–62.

Note that this guidance is all predicated on the use of large, intact, largely supercoiled plasmid DNA. There are specific risks noted:

risk of introducing plasmids with strong regulatory regions into the host genome far exceeds that associated with random point mutations

The SV40 enhancer/promoter is one of the strongest regulatory regions known to modern molecular biology, arguably exceeded by the commonly used corresponding cytomegalovirus (CMV) enhancer/promoter sequences. These SV40 sequences are the ones which the EMA falsely asserts are “considered to be a non-functional part of the structure of the source plasmid” and further asserts (in the absence of actual cited data or reference) that “There is no scientific evidence that any of these SV40 fragments can act as insertional mutagens.”.

the technology used to detect plasmid persistence does not examine the frequency with which short fragments of plasmid integrate. In this context, sections of DNA as short as 7 bp can affect rates of integration or recombination.

In other words, the technology used to establish integration and recombination risk and safety in DNA vaccines does not apply to the DNA fragments (technically referred to as “oligonucleotides”) present after being “broken down and removed to what EMA considers to be a safe and acceptable level” and delivered to cells and tissues using the highly efficient self-assembling lipid nanoparticle complexes designed by Dr. Pieter Cullis and colleagues. The ONLY way to determine whether these shorter DNA fragments (including the SV40-derived fragments) do or do not present a risk to those accepting mod-mRNA vaccines that are contaminated/adulterated with plasmid DNA fragments - many of which are quite small- is to actually perform the studies required to objectively assess that risk.

To do otherwise is to, once again, substitute hope for data.

“I think it was hope, that the vaccines would work in that way” Dr. Deborah Birx, congressional testimony

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US Food and Drug Administration

The following is the response provided to reporter Zack Stieber of The Epoch Times, and kindly shared with me ahead of his own article which should be published shortly.

The claim that the FDA is required to take any of the authorized or approved mRNA COVID-19 vaccines off the market is false. With over a billion doses of the mRNA vaccines administered, no safety concerns related to the sequence of, or amount of, residual DNA have been identified. With regard to the FDA-approved mRNA vaccines, available scientific evidence supports the conclusion that they are safe and effective.

Here is the analogous response provided to journalist Mary Beth Pfeiffer:

The claim that the FDA is required to take any of the authorized or approved mRNA COVID-19 vaccines off the market is false. With over a billion doses of the mRNA vaccines administered, no safety concerns related to the sequence of, or amount of, residual DNA have been identified. With regard to the FDA-approved mRNA vaccines, available scientific evidence supports the conclusion that they are safe and effective.

Additional questions may be directed to the manufacturer.

Commentary:

In other words, the FDA does not feel it necessary to provide even a modicum of data to support their position. Note that cancer, birth defects and other mutations can take years to clinically manifest, so the 14 day VAERS reporting window will provide no useful data to address these risks.

The FDA provides no documentation to support their (false) assertion that these products are “safe and effective”, and in fact that statement is basically more propaganda. No context is provided to support the entirely subjective terms “safe and effective”. Childhood seizures post administration, myopericarditis, coagulopathy, stroke, autoimmune diseases including Guillain–Barré syndrome (GBS) and finally reproductive harm including alteration of menstruation are all officially acknowledged risks. NOT safe by my subjective criteria. The products do not prevent infection, spread, hospitalization or death from COVID-19. That is also not in dispute. The FDA and CDC have been reduced to asserting that these products reduce the risk of hospitalization and death, but that has now become a highly controversial position due to the global evidence of negative effectiveness beginning at some period of months after administration. They are NOT effective by my subjective criteria. What are the specific criteria which the FDA is employing to suggest otherwise?

Show me both the criteria as well as the data to support these claims. By congressional statute, American citizens are owed proof of effectiveness, safety, purity, and lack of adulteration. This is the one job which congress has authorized for FDA to perform.

AS to the assertion that “no safety concerns related to the sequence of, or amount of, residual DNA have been identified”; this is directly refuted by the preceding text including the cited article describing the risks of short DNA fragments, and the risks of highly active regulatory sequences such as the SV40 enhancer/promoter. This statement is clearly false.

Until data are transparently provided for independent review demonstrating that the risk of integration associated with the allowed level of DNA fragments and the specific lipid nanoparticle delivery system employed for these mod-mRNA products is not clinically significant, any such statement by FDA should be regarded as disinformation provided for political purposes- ergo political propaganda.

Show American citizens the data to support these claims or they must conclude that such data do not exist.

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Health Canada

Inquiry:

From: Mary Beth Pfeiffer
Sent: Sunday, October 22, 2023 4:23 PM
To: HEALTH MEDIA SANTÉ (HC/SC)
Subject: Inquiry from US Journalist on SV40/DNA contamination

Hello,

I am writing an article that will include comments from vaccine experts who are concerned about the finding of Simian Virus 40 DNA in vials of mRNA Covid vaccine. 

Can you confirm the statement reported by the Epoch Times under the headline: Health Canada Confirms Undisclosed Presence of DNA Sequence in Pfizer Shot?

Can you also email me a copy of your statement or a link to it? 

Does this so-called "adulteration" constitute cause to remove the mRNA vaccines from the market?

I am on deadline so a speedy response is welcome. Your office has answered my queries in the past, for which I am grateful.

Thank you.

Mary Beth Pfeiffer
Investigative Journalist

Response:

Health Canada initially authorized the Pfizer-BioNTech COVID-19 mRNA vaccine in December 2020 and subsequently has authorized updated versions, including the most recent vaccine targeting the XBB Omicron subvariant in September 2023. Each assessment included a determination that the vaccine met the Department's stringent regulatory safety, efficacy and quality requirements for use in Canada.

As a regulator, Health Canada sets quality standards and requirements for manufacturers to follow, including providing comprehensive and detailed information about the vaccine itself, and about the manufacturing process. In the manufacture of any vaccine, residual elements that are part of the standard manufacturing process may remain. There are strict limits and controls for the presence of these residual fragments to ensure that there is no effect on the safety or effectiveness of the vaccine.     

The Pfizer-BioNTech COVID-19 vaccine does not contain simian virus 40 (SV40). The presence of the SV40 promoter enhancer sequence is not the same as the presence of the whole virus itself. 

The SV40 promoter enhancer sequence was found to be a residual DNA fragment in Pfizer-BioNTech COVID-19 vaccine. The fragment is inactive, has no functional role, and was measured to be consistently below the limit required by Health Canada and other international regulators.  

Any claims that the presence of the SV40 promoter enhancer sequence is linked to an increased risk of cancer are unfounded. There is also no evidence to support that the presence of the full SV40 in any vaccine increases the risk of cancer or the acceleration of cancer in individuals. 

Health Canada continues to monitor the COVID-19 vaccines to ensure that they continue to meet the highest standards for safety, effectiveness and quality and that their benefits continue to outweigh any potential risks.

Commentary:

All of these points have been covered above.

There are strict limits and controls for the presence of these residual fragments to ensure that there is no effect on the safety or effectiveness of the vaccine. 

What are those limits, how were they established, and what studies were performed to demonstrate that those limits ensure that there is no effect on the safety of the vaccine?

Show us the data.

The fragment is inactive, has no functional role

Any claims that the presence of the SV40 promoter enhancer sequence is linked to an increased risk of cancer are unfounded.

As summarized above, these statements are incorrect. These comments are merely unsubstantiated and unsupported propaganda. The SV40 sequences including an origin of DNA replication which is active in mammalian cells (that is why it is included in the plasmid!), as well as including a highly active genetic switch which is also functional in mammalian cells (the enhancer/promoter)- a strong regulatory region. As noted in the peer reviewed publications cited above, “risk of introducing plasmids with strong regulatory regions into the host genome far exceeds that associated with random point mutations “. … In this context, sections of DNA as short as 7 bp can affect rates of integration or recombination. Examples include the VDJ recombination signal sequence and related sequences, chi-like elements and minisatellites, ALU sequences, a recombinase signal present in hepatitis B and mammalian genomes, and topoisomerase II recognition sites.

Conclusion

“They lie to us, we know they're lying, they know we know they're lying, but they keep lying to us, and we keep pretending to believe them.” Attributed to Alexander Solzhenitsyn.

This substack essay has addressed the scientific data relating to the risks of plasmid DNA fragment contamination/adulteration of the mod-mRNA vaccines, for which there have been “over a billion doses of the mRNA vaccines administered” according to the FDA. Correspondence on this topic from the European Medicines Agency (EMA), US Food and Drug Administration (FDA) and Health Canada has been provided verbatim (“the receipts”) for reader review, and key statements made have been reviewed in light of existing peer reviewed scientific literature spanning decades. Unsupported assertions and blanket denials by these regulatory agencies concerning the biologic activity and potential risks of the SV40 origin-enhancer-promoter sequences have been highlighted and readily refuted (fact checked as disinformation). Key publications and regulatory guidance concerning integration and genotoxicity risk assessment authored by National Cancer Institute and FDA scientists have been cited and relevant text refuting false assertions made by these regulatory agencies have been highlighted.

In sum, this information clearly documents the shoddy, biased regulatory review processes employed by these agencies concerning the rushed mod-mRNA “vaccine” regulatory oversight and approval processes. There is a clear appearance of conflict of interest associated with current practices, which stand in stark contrast to the cautious and methodical assessments of integration and genotoxicity risks which characterized previous historic regulatory authority approaches to the closely related field of DNA vaccines. Risks previously clearly identified by regulatory authorities as associated with DNA vaccines, which inform risk assessment for the DNA fragments which all three regulatory authorities now admit are present in the mod-mRNA vaccines, have been casually overlooked and summarily denied despite a profound lack of data to support and justify these regulatory positions.

This analysis and information summary clearly documents a profound failure of all three regulatory authorities to do their most important job, to protect their citizens from risks of both inadequately tested and poorly controlled injectable products which are neither safe nor effective, and to prevent exposure of the public to adulterated pharmaceutical products. In stark contrast to the disinformation propaganda which is being offered by each of these regulatory authorities, the public deserves open, comprehensive and transparent communication of risk and/or the data demonstrating lack of risk associated with delivery of these DNA fragment (oligonucleotide) contaminant/adulterants to a wide range of cell, tissue, embryo and fetus via the most efficient non-viral systemic delivery system yet devised by man.

The curt and dismissive regulatory responses received from each of these regulatory authorities suggest that such data do not exist, and instead biased regulatory opinion has once again been substituted for definitive data. The public should demand that regulatory authorities show the actual data supporting their assertions of safety. Both current and future generations of human beings deserve no less.

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Relevant additional references

(a few of the many- there are an abundance of peer reviewed papers documenting recombination and insertional mutagenesis associated with short DNA fragments):

Biol Cell. 2010 Oct;102(10):561-80.

Reverse genetics in eukaryotes

Serge Hardy ¹, Vincent Legagneux, Yann Audic, Luc Paillard

Abstract

Reverse genetics consists in the modification of the activity of a target gene to analyse the phenotypic consequences. Four main approaches are used towards this goal and will be explained in this review. Two of them are centred on genome alterations. Mutations produced by random chemical or insertional mutagenesis can be screened to recover only mutants in a specific gene of interest. Alternatively, these alterations may be specifically targeted on a gene of interest by HR (homologous recombination). The other two approaches are centred on mRNA. RNA interference is a powerful method to reduce the level of gene products, while MO (morpholino) antisense oligonucleotides alter mRNA metabolism or translation. Some model species, such as Drosophila, are amenable to most of these approaches, whereas other model species are restricted to one of them. For example, in mice and yeasts, gene targeting by HR is prevalent, whereas in Xenopus and zebrafish MO oligonucleotides are mainly used. Genome-wide collections of mutants or inactivated models obtained in several species by these approaches have been made and will help decipher gene functions in the post-genomic era.

Methods Mol Biol. 2014:1114:115-42.

Triplex-mediated genome targeting and editing

Faisal Reza ¹, Peter M Glazer

Abstract

Genome targeting and editing in vitro and in vivo can be achieved through an interplay of exogenously introduced molecules and the induction of endogenous recombination machinery. The former includes a repertoire of sequence-specific binding molecules for targeted induction and appropriation of this machinery, such as by triplex-forming oligonucleotides (TFOs) or triplex-forming peptide nucleic acids (PNAs) and recombinagenic donor DNA, respectively. This versatile targeting and editing via recombination approach facilitates high-fidelity and low-off-target genome mutagenesis, repair, expression, and regulation. Herein, we describe the current state-of-the-art in triplex-mediated genome targeting and editing with a perspective towards potential translational and therapeutic applications. We detail several materials and methods for the design, delivery, and use of triplex-forming and recombinagenic molecules for mediating and introducing specific, heritable, and safe genomic modifications. Furthermore we denote some guidelines for endogenous genome targeting and editing site identification and techniques to test targeting and editing efficiency.

Nucleic Acids Res 1987 Nov 25;15(22):9379-95.

Differential regulation of viral and cellular genes in F9 mouse embryonal carcinoma cells

M J Sleigh ¹

Abstract

Expression of genes driven by the SV40 promoter/enhancer appears to be under net negative regulatory control in undifferentiated F9 cells, but not in their differentiated derivatives. In cells containing integrated copies of an SV40 promoter-driven marker gene, induction of differentiation by retinoic acid treatment produced a modest increase in transcription from the viral promoter. A much greater increase was observed when differentiated or undifferentiated cells were treated with the protein synthesis inhibitor, cycloheximide. If cycloheximide acts through removal of negative-regulatory molecule(s), then it is apparent that these molecules are present in both differentiated and undifferentiated cells, and that retinoic acid treatment removes only a portion of the total transcriptional repression. RNA levels from a variety of cellular genes activated during F9 cell differentiation were either unaffected or only slightly increased by cycloheximide treatment. This suggests important qualitative or quantitative differences in the regulation mechanism for viral and cellular genes in differentiating F9 cells.

PLoS One 2014 Jan 21;9(1):e86324.

Novel therapeutic approaches for various cancer types using a modified sleeping beauty-based gene delivery system

In-Sun Hong ¹, Hwa-Yong Lee ¹, Hyun-Pyo Kim ²

Abstract

Successful gene therapy largely depends on the selective introduction of therapeutic genes into the appropriate target cancer cells. One of the most effective and promising approaches for targeting tumor tissue during gene delivery is the use of viral vectors, which allow for high efficiency gene delivery. However, the use of viral vectors is not without risks and safety concerns, such as toxicities, a host immune response towards the viral antigens or potential viral recombination into the host's chromosome; these risks limit the clinical application of viral vectors. The Sleeping Beauty (SB) transposon-based system is an attractive, non-viral alternative to viral delivery systems. SB may be less immunogenic than the viral vector system due to its lack of viral sequences. The SB-based gene delivery system can stably integrate into the host cell genome to produce the therapeutic gene product over the lifetime of a cell. However, when compared to viral vectors, the non-viral SB-based gene delivery system still has limited therapeutic efficacy due to the lack of long-lasting gene expression potential and tumor cell specific gene transfer ability. These limitations could be overcome by modifying the SB system through the introduction of the hTERT promoter and the SV40 enhancer. In this study, a modified SB delivery system, under control of the hTERT promoter in conjunction with the SV40 enhancer, was able to successfully transfer the suicide gene (HSV-TK) into multiple types of cancer cells. The modified SB transfected cancer cells exhibited a significantly increased cancer cell specific death rate. These data suggest that our modified SB-based gene delivery system can be used as a safe and efficient tool for cancer cell specific therapeutic gene transfer and stable long-term expression.

Nat Rev Drug Discov 2020 Oct;19(10):673-694.

Advances in oligonucleotide drug delivery

Thomas C Roberts ^1 2, Robert Langer ³, Matthew J A Wood ^4 5

Abstract

Oligonucleotides can be used to modulate gene expression via a range of processes including RNAi, target degradation by RNase H-mediated cleavage, splicing modulation, non-coding RNA inhibition, gene activation and programmed gene editing. As such, these molecules have potential therapeutic applications for myriad indications, with several oligonucleotide drugs recently gaining approval. However, despite recent technological advances, achieving efficient oligonucleotide delivery, particularly to extrahepatic tissues, remains a major translational limitation. Here, we provide an overview of oligonucleotide-based drug platforms, focusing on key approaches - including chemical modification, bioconjugation and the use of nanocarriers - which aim to address the delivery challenge.

Pieter Cullis and his group are the ones that developed the self assembling lipid nanoparticle technology used for all of the mod-mRNA vaccines

Nat Nanotechnol. 2021 Jun;16(6):630-643.

The current landscape of nucleic acid therapeutics

Jayesh A Kulkarni ^# 1 2 3, Dominik Witzigmann ^# 2 3 4, Sarah B Thomson ¹, Sam Chen ⁵, Blair R Leavitt ¹, Pieter R Cullis ^2 4, Roy van der Meel 

Abstract

The increasing number of approved nucleic acid therapeutics demonstrates the potential to treat diseases by targeting their genetic blueprints in vivo. Conventional treatments generally induce therapeutic effects that are transient because they target proteins rather than underlying causes. In contrast, nucleic acid therapeutics can achieve long-lasting or even curative effects via gene inhibition, addition, replacement or editing. Their clinical translation, however, depends on delivery technologies that improve stability, facilitate internalization and increase target affinity. Here, we review four platform technologies that have enabled the clinical translation of nucleic acid therapeutics: antisense oligonucleotides, ligand-modified small interfering RNA conjugates, lipid nanoparticles and adeno-associated virus vectors. For each platform, we discuss the current state-of-the-art clinical approaches, explain the rationale behind its development, highlight technological aspects that facilitated clinical translation and provide an example of a clinically relevant genetic drug. In addition, we discuss how these technologies enable the development of cutting-edge genetic drugs, such as tissue-specific nucleic acid bioconjugates, messenger RNA and gene-editing therapeutics.

Nucleic Acids Res. 1993 Feb 11;21(3):407-12.

Targeted recombination with single-stranded DNA vectors in mammalian cells

K Fujioka ¹, Y Aratani, K Kusano, H Koyama

Abstract

We studied the ability of single-stranded DNA (ssDNA) to participate in targeted recombination in mammalian cells. A 5' end-deleted adenine phosphoribosyltransferase (aprt) gene was subcloned into M13 vector, and the resulting ssDNA and its double-stranded DNA (dsDNA) were transfected to APRT-Chinese hamster ovary cells with a deleted aprt gene. APRT+ recombinants with the ssDNA was obtained at a frequency of 3 x 10(-7) per survivor, which was almost equal to that with the double-stranded equivalent. Analysis of the genome in recombinant clones produced by ssDNA revealed that 12 of 14 clones resulted from correction of the deletion in the aprt locus. On the other hand, the locus of the remaining 2 was not corrected; instead, the 5' deletion of the vector was corrected by end extension, followed by integration into random sites of the genome. To exclude the possibility that input ssDNA was converted into its duplex form before participating in a recombination reaction, we compared the frequency of extrachromosomal recombination between noncomplementary ssDNAs, and between one ssDNA and one dsDNA, of two phage vectors. The frequency with the ssDNAs was 0.4 x 10(-5), being 10-fold lower than that observed with the ssDNA and the dsDNA, suggesting that as little as 10% of the transfected ssDNA was converted into duplex forms before the recombination event, hence 90% remained unchanged as single-stranded molecules. Nevertheless, the above finding that ssDNA was as efficient as dsDNA in targeted recombination suggests that ssDNA itself is able to participate directly in targeted recombination reactions in mammalian cells.

Mol Cell Biol 1996 Jan;16(1):10-8.

High-frequency illegitimate integration of transfected DNA at preintegrated target sites in a mammalian genome

R V Merrihew ¹, K Marburger, S L Pennington, D B Roth, J H Wilson

Abstract

To examine the mechanisms of recombination governing the illegitimate integration of transfected DNA into a mammalian genome, we developed a cell system that selects for integration events in defined genomic regions. Cell lines with chromosomal copies of the 3' portion of the adenine phosphoribosyltransferase (APRT) gene (targets) were established. The 5' portion of the APRT gene, which has no homology to the integrated 3' portion, was then electroporated into the target cell lines, and selection for APRT gene function was applied. The reconstruction of the APRT gene was detected at frequencies ranging from less than 10(-7) to 10(-6) per electroporated cell. Twenty-seven junction sequences between the integrated 5' APRT and its chromosomal target were analyzed. They were found to be randomly distributed in a 2-kb region immediately in front of the 3' portion of the APRT gene. The junctions fell into two main classes: those with short homologies (microhomologies) and those with inserted DNA of uncertain origin. Three long inserts were shown to preexist elsewhere in the genome. Reconstructed cell lines were analyzed for rearrangements at the target site by Southern blotting; a variety of simple and complex rearrangements were detected. Similar analysis of individual clones of the parental cell lines revealed analogous types of rearrangement, indicating that the target sites are unstable. Given the high frequency of integration events at these sites, we speculate that transfected DNA may preferentially integrate at unstable mammalian loci. The results are discussed in relation to possible mechanisms of DNA integration.

Nature. 2014 Oct 16;514(7522):380-4.

CRISPR-mediated direct mutation of cancer genes in the mouse liver

Wen Xue ¹, Sidi Chen ¹, Hao Yin ¹, Tuomas Tammela ², Thales Papagiannakopoulos ², Nikhil S Joshi ², Wenxin Cai ², Gillian Yang ², Roderick Bronson ³, Denise G Crowley ², Feng Zhang ⁴, Daniel G Anderson ⁵, Phillip A Sharp ⁶, Tyler Jacks ⁷

Abstract

The study of cancer genes in mouse models has traditionally relied on genetically-engineered strains made via transgenesis or gene targeting in embryonic stem cells. Here we describe a new method of cancer model generation using the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system in vivo in wild-type mice. We used hydrodynamic injection to deliver a CRISPR plasmid DNA expressing Cas9 and single guide RNAs (sgRNAs) to the liver that directly target the tumour suppressor genes Pten (ref. 5) and p53 (also known as TP53 and Trp53) (ref. 6), alone and in combination. CRISPR-mediated Pten mutation led to elevated Akt phosphorylation and lipid accumulation in hepatocytes, phenocopying the effects of deletion of the gene using Cre-LoxP technology. Simultaneous targeting of Pten and p53 induced liver tumours that mimicked those caused by Cre-loxP-mediated deletion of Pten and p53. DNA sequencing of liver and tumour tissue revealed insertion or deletion mutations of the tumour suppressor genes, including bi-allelic mutations of both Pten and p53 in tumours. Furthermore, co-injection of Cas9 plasmids harbouring sgRNAs targeting the β-catenin gene and a single-stranded DNA oligonucleotide donor carrying activating point mutations led to the generation of hepatocytes with nuclear localization of β-catenin. This study demonstrates the feasibility of direct mutation of tumour suppressor genes and oncogenes in the liver using the CRISPR/Cas system, which presents a new avenue for rapid development of liver cancer models and functional genomics.