Last week, I discovered a new antivax scientist named Kevin McKernan, whose message had been recently amplified by long time quack tycoon Joe Mercola. At the time, his false claim was that SV40 promoter sequences in plasmid DNA contaminating the Pfizer and Moderna mRNA-based COVID-19 vaccines were somehow putting people at risk for cancer. In my deconstruction, I pointed out that this claim was an even more ridiculous variation on an old antivax claim that oral polio vaccines from the late 1950s to 1963 that had been discovered to be contaminated with SV40 virus were responsible for a wave of cancer. They weren’t, but this antivax claim keeps coming back from the grave (going back a decade, at least), ready to party. This time is no different, except that the claim is even more ridiculous than the movie from which I adapted that tagline to describe it. Why? Because it’s almost as though McKernan, his background in microbiology and (apparently) genomics, either does not know the difference between a promoter sequence and an actual gene or, more likely, knows the difference but knows that his audience doesn’t know the difference but is aware of the polio vaccine/SV40 story.(Take your pick.)
This time around, McKernan is back on his Substack with what he views, apparently, as a refutation of the most common refutations of the antivax claim that COVID-19 vaccines, being mRNA vaccines, get into the nucleus and “permanently alter your DNA.” By way of background, I’ve long been discussing how the idea that COVID-19 vaccines will “damage” or somehow “irreversibly alter” your DNA started showing up months before the mRNA-based vaccines even won emergency use approval and began to be distributed, and I’ve been pointing out how such a claim bespeaks an utter ignorance and/or misunderstanding of some very basic molecular biology of the sort that is taught in introductory level biology classes in college. None of this has stopped antivaxxers from citing studies that are claimed to demonstrate that the mRNA from the vaccine can somehow reverse transcribe itself and then integrate its gene sequence for the COVID-19 spike protein into the DNA of the cell’s nucleus but, when examined by people with molecular biology expertise, are fairly trivially shown not to be good evidence of anything of the sort. Basically, there’s no evidence that the mRNA from the vaccine gets into the nucleus or is reverse transcribed into DNA, much less “permanently alters” your DNA. Even if it could, also remember that the cells that primarily take up the lipid nanoparticles of the vaccine containing the mRNA for the spike protein are muscle cells, which do not divide.
Which brings me to McKernan’s claim, embodied in the title of his post, Nuclear permeability during cell division, which he bills as “Flattening the ‘won’t get to the nucleus’ canard.” He begins with a whole lot of handwaving about his previous claims:
First, we need to do some numbers.
The 100ug Moderna shots deliver over 40 trillion mRNA molecules (14T for Pfizer) and the number of LNPs per shot is reported to be 10-50 billion. For easy math, lets assume 40 billion LNPs. This is roughly 1,000 mRNAs in each LNP.
What most people who don’t do molecular biology don’t know that McKernan does know is that 40 trillion molecules is not a huge number. One mole is 6.023 x 1023 molecules, which makes 10 trillion molecules 1.66 x 10-14, or 16.6 femtomole. Of course, McKernan transparently chose the Moderna vaccine as his example because it has a lot more mRNA in it (100 μg as opposed to 30 μg), resulting in 10 trillion, a bigger number than 3 trillion. Either way, this is not a huge number of molecules, as huge a number as it seems. McKernan is doing nothing more than pulling a common trick antivaxxers like to use to make the vaccines seem scarier. Look at how many molecules of the scary spike protein mRNA they contain!
Riffing on this, he then fear mongers about the modified uracil base that is used to stabilize the mRNA in the vaccines because native mRNA is very unstable and degrades easily:
There over 800 N1-methyl-PseudoUs (m1Ψ) in each mRNA of Pfizer and likely an equal number in Moderna mRNAs for C19 vaccination. 800,000 m1Ψ per cell. This may influence the PUS (Pseudo Uridine Synthase) pathway. This pathway is very thinly published on. This is a very nascent field in the Epigenetics of RNAs. While we understand which enzyme modify Uracil to Ψ to m1Ψ , less is known about the processes that reverse this.
Most other RNA methylation systems (methyl A, and methyl C) have RWE pathways (Read, Write, Erase) understood. There are proteins identified that recognize the modified base (read), ones that make the modifications (write), and proteins identified that erase the methylation. The reading and writing for Ψ and m1Ψ have been identified but the erasing is still a mystery.
Of course, this is nothing more than a fear mongering appeal to ignorance. In fact, quite a lot is known about using this form of uracil in mRNAs used to express various proteins in cells, but McKernan pulls the claim that this modified base might influence the PUS pathway, all while invoking a newer field, the epigenetic of mRNA processing, in much the way that quacks have long been deceptively invoking epigenetic modifications of DNA. (Deepak Chopra is notorious for invoking “epigenetics” to “explain” how emotions can supposedly cure disease. Indeed, a decade ago I quipped that “epigenetics” had become the new “quantum” to quacks.) So is this something to worry about? If it is, certainly he presents no evidence, although PUS enzymes are a fascinating topic whose study could well result in a better understanding of RNA transcription.
Next up, McKernan invokes the dreaded DNA contamination, much as pathologist Sin Han Lee did a decade ago when he used an incredibly sensitive nested PCR assay prone to false positives to “prove” that there was “fetal DNA” in Gardasil, even though what he discovered, if legit, was still such an incredible small quantity of DNA left over from the cell line used in the manufacture of the vaccine as to be harmless. Here we go with double-stranded DNA (dsDNA):
If dsDNA contamination is 1%-10% of these mRNA numbers we are injecting 10-100 dsDNA molecules into each cell transfected. Likely 40 billion cells if 1 LNP infects a single cell. In reality this number is likely much higher at the site of the injection and diffuses to lower MOI (multiplicity of infection) as it distributes to nearly every tissue in the body. This is 0.13% of the 30 trillion cells in your body.
Cells are being transfected with 10-100 dsDNA molecules each. These DNAs are likely fragmented and short dsDNA given what we know with the low Adverse event batches analyzed to date. They may be longer and more intact in the poor batches analyzed by Schmeling et al.
Will the dsDNA contaminants ever make it to the nuclease? About 1/40th of them will contain an SV40 promoter with a nuclear localization signal. The dsDNA isn’t all SV40 DNA. It’s 7,810bp of sequence for Pfizer. If the dsDNA is ~200bp in size The SV40 region is just ~1/40th of the dsDNA in the contamination.
Notice again how McKernan plays with large numbers in order to make the vaccine sound scary. That first part is nothing more than an acknowledgment that the cells near the injection site likely take up many lipid nanoparticles (LNPs) each, while cells much further away take up many fewer. Of course, “multiplicity of infection” is a word commonly used to describe viral infection using viruses as vectors for introducing genes into cells and generally describes how many virus particles on average infect each cell in a given experiment. When we do in vitro experiments (as I’ve done many times with lentivirus vectors), we frequently calculate the MOI to make sure that we have enough virus to make sure that multiple copies of the virus infect each cell in the dish; we also use MOI for dose-response studies in which the amount of protein product of interest made by the virus is plotted against MOI. Uptake of LNPs by cells is not an infectious process but rather what we in the biz call a transfection, and the way we normally describe this is as a concentration or mass of transfecting agent per cell, such as this paper looking at lipid nanoparticle drugs for breast cancer. No doubt McKernan knows all this but chooses MOI because it uses the word “infection” and thus sounds scarier to lay people, even though the mRNA-based COVID-19 vaccines do not “infect” anything.
Also notice how McKernan admits something that I pointed out the last time I discussed his fear mongering about plasmid DNA “contamination” of mRNA vaccines, namely that the DNA fragments are almost certainly all short fragments, which means that they can’t make any proteins. It is true that the SV40 promoter is only 72 base pairs (bp) long, but, again, it doesn’t code from anything, and even if it does have a nuclear localization signal; i.e., a short sequence that leads a DNA molecule to be taken to the nucleus from the cytoplasm. Again, he’s just handwaving. Even in his paper touted by Mercola, he never actually showed that any of the contaminating plasmid DNA that he claimed to have found actually found its way into the nucleus of any cells, which is where it would have to get in order to do anything.
Don’t worry, though. He has another way to suggest without proving that vaccines “permanently alter your DNA”:
But Dr. Bhakdi pointed out something I hadn’t considered. During cell division the nucleus disassembles exposing the nuclear genome to the cytoplasm and transcription is still active during this time window. Most genome integrations occur in regions being actively transcribed. This would imply all fragments of the 7,810bp plasmid DNA is on the table for integration.
McKernan forgets to consider—or, more likely, didn’t forget but simply declined to mention—is the cells targeted by the LNPs carrying the vaccine’s mRNA. Think about it. Where is the vaccine injected? That’s right: Into muscle. Now, think about it. Do muscle cells divide? No, they do not. They are what is referred to as terminally differentiated. While the idea that terminally differentiated cells do not and cannot ever divide again is a bit simplistic, it is true that normally muscle cells do not divide under normal circumstances. Indeed, because muscle cells are terminally differentiated, a lot of research has gone into trying to “reactivate” them into muscle precursor cells that can divide.
What this means is that, in practice, it doesn’t matter if dividing cells have higher nuclear membrane permeability and thus allow the entry of mRNA and fragments of DNA from plasmids into their nucleus, where the vaccine can “permanently alter your DNA.” It’s irrelevant. Of course, antivaxxers like to point to biodistribution studies showing that the LNPs go elsewhere, something that I have discussed in detail before, pointing out that huge doses of LNPs were given to rats to determine these results. The idea of these experiments is to detect any organ that any amount of LNP might deposit in, no matter how small the amount. Antivaxxers weaponized the observation that a small amount of such huge doses deposited in the ovaries to claim that the vaccines cause infertility, even though they do not. I note that, even looking at the chart, the tissues to which the LNPs go are all for the most part either terminally differentiated or, even if capable of division, mostly in a quiescent, not dividing unless injury requires repair and regeneration. (The endothelial cells lining blood vessels are a good example; they are quiescent—not dividing—most of the time but can begin dividing in response to injury in order to repair damage.)
Unsurprisingly, McKernan also appears to have anticipated the argument that he’s fear mongering about tiny amounts of mRNA and DNA that are harmless at the amounts used:
Many of the detractors to this work will Namaste over the dsDNA being too little to matter. Remind them of using qPCR to detect C19 sgRNA from your mucosa. They were calling some positive for CTs under 40.
Violating civil rights and wrecking the economy came with far lower standards than the contaminants of their pharmaceutical partners. The dsDNA contaminant in these vaccines is a CT of 20 for 1/300th of the vaccine dose. Thats over a millions times higher (300 million for a CT of 40) amounts of contaminating nucleic acid than what would trigger a CT 40 sgRNA qPCR test.
With qPCR, we finally have a quantitative Political Corruption Readout as we can measure precisely the size of their hollowed out house of hypocrisy.
This harkens back to an old conspiracy theory about the PCR tests used to diagnose COVID-19 and the CT (cycle threshold) used as “positive.” The long version of the explanation of this conspiracy theory is here. The CliffsNotes version follows.
Anyone who’s done PCR (polymerase chain reaction), a test that uses small sequences from a known DNA sequence to amplify it, knows that cranking up the number of cycles does indeed increase the chances of amplifying a contaminant or a sequence that is similar, but not identical, to the intended target sequence. Indeed, setting the threshold count (CT) cutoff on a PCR test is a balancing act, in which you balance the increased sensitivity of using a higher CT and more amplification cycles as your cutoff and the attendant possibility of more false positives associated with that more sensitive cutoff against the possibility of missing a lot more cases of real infection if you use a lower CT. Back in the day, the “casedemic” conspiracy theory claimed that setting the CT at 40 cycles was too high and therefore produced a lot of false positives, making most “positive” PCR tests false positives. They weren’t. Yes, false positives were a problem, just not nearly as big a problem as COVID-19 conspiracy theorists claimed and certainly not an indication that COVID-19 numbers were being vastly inflated for nefarious purposes, as the “casedemic” conspiracy theory claimed.
McKernan also knows that this is a false comparison, which is likely why he pivots to reproducing chapters from an old textbook by Robert Weinberg on “Ocogenesis”—sorry, I couldn’t resist the spelling burn over “oncogenesis,” or the development of cancer—to try to convince you that the amounts of DNA fragments, which are tiny even if his inflated estimates of how much is in the vaccine are accurate, can cause cancer. Anyone who knows anything about molecular biology and cancer will likely laugh at this. (I did.) The book pages reproduced discuss oncogenic retroviruses and have nothing to do with what McKernan says:
A few excerpts from Robert Weinberg’s text book on Ocogenesis were sent to me from some colleagues in Japan. This is a reminder of the carcinogenic risks of genome integration which is far harder for your body to clean up when these injections also lower your white blood cell counts.
Again, read the pages. Even without a knowledge of molecular biology, you can see that the pages are talking about retroviruses, namely RNA viruses that can be reverse transcribed into DNA, which can then integrate into the genome. Retroviruses (like, for instance, HIV) are a very different thing from mRNA in LNPs or even short fragments of plasmid DNA, neither of which have the ability to replicate in cells. The mRNA from the vaccine doesn’t have the ability to reverse transcribe itself into DNA that can be integrated into the host cell’s genome. The minuscule number of DNA fragments can’t integrate into the host genome either. (At least, if they can, certainly McKernan has not demonstrated it or that it causes any harm, particularly in terminally differentiated cells.)
Of course, McKernan is somewhat more sophisticated than that. What he seeks to claim is that insertional mutagenesis. Let me quote the relevant passage from the Weinberg text:
Suddenly, all the clues needed to solve the puzzle of leukemogenesis (leukemia formation) by ALV fell into place. The solution went like this. During the course of infecting a chicken, ALV spread to thousands, then millions of cells in the hematopoietic system of this bird. Soon, the infection was so successful that the bird would become viremic, that is, its bloodstream carried high concentrations of virus particles. Each of these tens of millions of infections resulted in the insertion of an ALV provirus at some random location in the genome of an infected cell. In the vast majority of cases, this provirus integration had no effect on the infected host cell, aside from forcing the host to produce large numbers of progeny virus particles. But on rare occasions, perhaps in 1 out of 10 million infections, a provirus became integrated by chance next to the c-myc gene (Figure 3.23B). This jackpot event led to an explosive outcome—conversion of the c-myc gene into a potent oncogene whose unceasing expression was now driven by the adjacently integrated provirus and its transcriptional promoter. The rare cell carrying this deregulated myc gene then began uncontrolled proliferation, and within weeks, some of the progeny cells evolved further into more aggressive cancer cells that constituted a leukemia.
This scenario explains the slow kinetics with which these leukemias arise after initial viral infection of a bird. Since activation of the c-myc gene through provirus integration is a low-probability event, many weeks and many millions of infectious events are required before these malignancies are triggered. This particular mechanism of protooncogene activation came to be called insertional mutagenesis; it explains, as well, the leukemogenic powers of other slowly acting retroviruses, such as MLV. By now, study of avian and murine retrovirus-induced infections has demonstrated integration events next to more than 25 distinct cellular proto-oncogenes. Indeed, insertional mutagenesis can be used as a powerful strategy to find new proto-oncogenes…
I trust that most readers can see how an infection with a virus that is actively replicating and producing so many copies of itself in an organism that low probability events become relevant is different from LNPs containing mRNA and maybe a tiny amount of contaminating plasmid DNA fragments. From a molecular biology standpoint, McKernan’s argument is profoundly dishonest. Yes it is possible that DNA fragments that get into cells can integrate in the genome, but the likelihood is very small and orders of magnitude smaller that any oncogenes will be activated. In nonreplicating cells, it wouldn’t even matter if oncogenes were activated by insertional mutation because, again, these cells don’t replicated and replication is required for carcinogenesis. Why do you think the virus described above caused leukemia and not muscle tumors? Bone marrow cells replicate; muscle does not. Also, if McKernan’s fear mongering about insertional oncogenesis were valid, we’d expect to have seen myosarcomas (sarcomas arising from muscle cells) at vaccine injection sites. (After all, that’s where the vast majority of the vaccine ends up and stays.) We have not.
The bottom line is that Kevin McKernan is using his knowledge of genomics to frighting people about the mRNA-based COVID-19 vaccines. What he is claiming, while not as implausible as homeopathy, is pretty damned implausible, and he shows no evidence that any of the mechanisms that he’s trying to scare you with are operative with respect to the Pfizer or Moderna COVID-19 vaccines, other than that he’s detected small amounts of plasmid DNA contamination, likely leftover from the manufacturing process. He has not shown that this DNA can get into the nucleus, much less integrate into the genome and “permanently alter your DNA.” His invocation of increased permeability of the nuclear membrane during cell division is nothing more than more handwaving, particularly given that the vast majority of the cells that take up the LNPs do not replicate. While I like to think that people like McKernan are speaking out of ignorance rather than deception (i.e., spreading misinformation rather than disinformation), McKernan should know better, given his training and expertise. Make of that observation what you will.