Integration into the genome... yet. They need long read Whole genome sequencing to prove that which Im sure is currently underway. Rudi showed it was integrated but was critiqued that it could be an artifact of making sequencing libraries. This paper fills in many questions.
The study was well controlled with RNAse confirmation that the target is in fact DNA and not RT of RNA.
Team "Scientific Censorship" has a lot of explaining to do. Many people on this team were vocal opponents of the CRISPR baby experiments run in China. Now they are guilty of
Advocating such experiments on a billion people. It is amazing how pliable their ethics are with a pinch of fear.
We covered this in our PrePrint with Dr. McCullough. @P_McCulloughMD
In a sane world this would lead to immediate moratorium on mRNA transfections until WGS are complete.
We are no longer in a sane world and you should take protection into your own hands osf.io/bcsa6/
You can find them at the below link on the bioRxiv preprint.
A good question from a fellow cat...
When will Whole Genome Sequence Moon?
I think they will need long read sequencing to sort out LINE-1 integration. That usually requires >100ng of DNA.
Human genome is 6pg so 10,000-100,000 cells of DNA are needed to sequence.
Each cell is unlikely to carry integrations in the same location of the genome. So they can't use consensus sequencing (ensemble of all 10,000 cells DNA sequencing) to sort it out. PacBio HiFi sequencing can deliver 150Gb in a single flow cell or 50X coverage per flow cell
More recent studies are pushing this number down to 5ng for PacBio, so they maybe able to get away with 1,000 cells. pacb.com/wp-content/upl…
This might require multiple flow cells. 10 would give you 500X. Either way, this could be over $25K in sequencing but the accuracy of the HiFi reads means each 20Kb read is as good as Sanger sequencing but 30X longer and better for mapping in LINE-1 regions.
They improve at a faster than Moore's law rate and they may be able to better speak to current yields. Will need to be done in those Huh7 cell lines and ultimate in patient biopsies. @PacBio pacb.com/wp-content/upl…
I did leave a comment in Rudi preprint that suggested DNAse and RNAse controls.
It had some push back.
The DNAse and RNAse experiments we just done on BNT162b2 and further move this from conspiracy towards Fact.
Before we get overly focused on Integration….
We should pause to realize this paper has data that should concern us today.
The paper demonstrates vax induced elevated LINE-1 expression.
It is worth reading the Back and Forth on the Jaenisch labs paper (Zhang et al).
They suggest integration with the virus is 2-5 integrations per 10,000 cells.
This is a dense read but it will help you to understand why the DNA fluorometer data is lighting up so much on modRNA. This is much more cross-talk than you see with traditional ssRNA.
That is because these are intercalating dyes that bind to minor grooves are only found in double stranded nucleic acids.
The fact that RNaseA is showing such a reduction in signal is because the RNA isnt single stranded.
At least half of it is digestable by a dsRNA specific nuclease known as RNaseIII.
Another portion responds to RNaseH which only digests RNA in RNA:DNA hybrids.
The fraction that responds to RNAseIII should worry you. Your cells have RNaseIII and its the start of the RNA interference pathway.
To make matters worse, Pharma is using an ELISA that is blind to dsRNA <400bp. It cant see this problem. And they know it exists
First…
They measure how long is spike expressed in cells.
7 days out it is higher than day 1. Peaks at day 5.
Using 3 different Fluorometry methods they zero in on DNA levels before and after RNaseA treatments.
4-5X over once RNA is removed.
The AccuBlue dyes had the least cross talk.
Your response to peoples request for your qPCR protocol is misinformation.
No transparency on your methods but we can already see that you are using only 1 assay in the vector (Kan gene) so you are under estimating the load 100X.
You should know this by now as Speicher et al published this a year ago.
@TGAgovau This is misinformation. Dr. David Speicher's latest test used RNaseA. There is no RNA interfering with the fluorometry.
Did you not even read the report?
Life must be good at the TGA when you can be a year behind all the time.
@TGAgovau Part of GMP and ISO is having your protocol open for inspection.
Is SV40 Large Tumor Antigen required for SV40 origin of replication activity.
No
Here we detect Pfizer plasmid DNA in a colon tumor biopsy 1 year after vaccination.
And its not small amounts of DNA. Its so much DNA that it can only be explained by plasmid amplification post vaccination or genome integration and amplification.
Variants are found in the SV40 Promoter that do not exist when you sequence the vaccine directly suggestive of replication errors once transfected into mammalian cells.