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Kevin McKernan

@Kevin_McKernan

Apr 14 • 4 tweets • 3 min read Twitter logo

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@Jikkyleaks

RNA-DNA hybrids.
What are they and how do they impact vaccine manufacturing and human health?
#plasmidgate
@Jikkyleaks @JesslovesMJK @ClownBasket

It turns out complementary N1-methylpseudouridine RNA inhibits DNAse I activity on contaminating plasmids.
This is likely the reason the vaccines are contaminated and the manufacturers should investigate other single stranded nucleases to clean this up.
anandamide.substack.com/p/dna-rna-hybr…

Be a real bummer if the antibiotic you used to grow your plasmid also inhibited the enzyme you planned to use to get rid of the plasmid.

pubmed.ncbi.nlm.nih.gov/10819299/

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More from @Kevin_McKernan

Kevin McKernan

@Kevin_McKernan

Apr 15

This is misunderstanding of polymerase fidelity.

The moment the RNA gets into the cell, it begins the process of making sgRNA and behaving just like the virus.

All of the ‘cloud’ issues begin at the first copy of the DNA->RNA and APOBEC response.

Likewise, making RNA from the DNA IC is NOT perfection error free replication. I’ve given him this literature?

DNA polymerases that replicate plasmids make an error every million bases. RNA polymerases that transcribe them make 10-100X more error.

So out of the gate he’s wrong on the fidelity topic and anyone can google the error rates of these enzymes.

book.bionumbers.org/what-is-the-er…

Read 12 tweets

Kevin McKernan

@Kevin_McKernan

Apr 12

The Pfizer Monovalent vaccine sequence is now public.
It also contains the 2 copy 72bp Enhancer in the SV40 promoter.

Another group in Europe sequenced this before us and shared their data.
They found the same thing using different methods.

More about it here-

open.substack.com/pub/anandamide…

The SV40 virus seen in the polio vaccines was the full 5kb virus.

We only have the promoter on these vectors.

We have no evidence they are carcinogenic. I do have concerns over SV40 promoters integrating into the genome near an oncogene.
Keith Peden paper is a good read on this

Read 4 tweets

Kevin McKernan

@Kevin_McKernan

Apr 11

This is an important paper.
One of the reasons we’ve been having a hard time quantitation the DNA in these vaccines is that the presence of the RNA stalls DNAse I and T5 exo.
That implies there are DNA/RNA hybrids or R-Loops in the vaccines.

nature.com/articles/s4158…

If you RNAse A treat the sample then the DNA is easily destroyed by nucleases.

#plasmidgate
.@Jikkyleaks @JesslovesMJK @sasha_latypova

Read 4 tweets

Kevin McKernan

@Kevin_McKernan

Apr 4

https://twitter.com/JesslovesMJK/status/1643128328298303488

It’s a very good presentation.
We are still trying to get a handle on how much of this is circular plasmid Vs fragments.
We have conflicting data on this that has prevented us from making quantitative statements.

1)our transformation efficiency is very low

https://twitter.com/JesslovesMJK/status/1643128328298303488

2)we don’t have sequence confirmation of those ecoli colonies.
3)we don’t have an electroporator. Heat shock is not the most efficient means.
3)we see a large peak on the Agilent gel that could be the linear or circular plasmid or…

4)there could be cross talk with the intercalating dyes on the gels where their DNA specificity is promiscuous with N1 methylpseudoU mRNA and the peak we think is a plasmid is just mRNA getting mis-stained on the DNA gels.

Read 8 tweets

Kevin McKernan

@Kevin_McKernan

Mar 30

H/T Dr. David Wiseman.

fda.gov/vaccines-blood…

Note the limit set by VRBPAC.
Note the differentiation of DNA types.
Protooncogenes…
Where does an SV40 promoter with a nuclear localization signal fit?