What is going on with the magnesium levels? Looked at the EMA document and nothing specific was found. Bet the FDA knows all this. Plus it explains the EMA asking for further work on the DNAse 1.
Also, I have always wondered why BioNTech didn't use its new nifty cellulose based chromatography method which gets rid of 90% of the dsRNA? Maybe it wasn't enough or not enough RNA yield.
The magnesium issue has not been considered yet, and I am currently at the stage of monitoring the verification since I'm currently busy with my day job and don't have enough time….
Thank you for presenting the paper. Katalin Karikó, BionTech, is one of the authors. I had a question in my mind about what dsRNA really is. There were some interesting points about the dsRNA production in this paper. That's the part below.
------
Phage RNA polymerases, including T7 RNA polymerase (T7RNAPol), transcribe the RNA with high fidelity from a DNA template containing the corresponding promoter. However, during the initiation of transcription, 5- to 11-nt-long RNA by-prod- ucts are generated as the enzyme aborts the synthesis with a certain probability.3,4 Considering that the T7RNAPol also has an obscure RNA-dependent as well as template-independent RNA polymerase activity,5–12 the short abortive RNA fragments and the 30 end of the full-length RNA can prime complementary RNA synthesis from the primary transcripts that leads to the generation of double-stranded RNA (dsRNA) contaminant. Recently, a promoter-independent transcription of full-length anti-sense RNA has been also reported as a novel mechanism of dsRNA generation in T7RNAPol-driven IVT reaction.13
The nuclei of human cells contain dsRNA that plays a role in nat- ural biological processes;14,15 however, when dsRNA enters the cells, from the extracellular milieu into their endosome or cytoplasm, it is sensed as a viral invader. All cells are capable of responding to dsRNA through sensors and effectors. Activation of dsRNA-depen- dent enzymes, such as oligoadenylate synthetase (OAS), RNA- specific adenosine deaminase (ADAR), and RNA-activated protein kinase (PKR), results in the inhibition of protein synthesis. In addi- tion, stimulation of the dsRNA sensors, e.g., Toll-like receptor 3 (TLR3), retinoic acid-inducible gene I (RIG-I), and melanoma dif- ferentiation-associated protein 5 (MDA5), leads to the secretion of different cytokines, including type I interferons, interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-a) (reviewed by Hart- mann16). Therefore, the elimination of dsRNA from the IVT mRNA is crucial to improve translation of the administered mRNA and to limit induction of cytokines, whether the mRNA products are intended for in vitro, ex vivo, or in vivo applications and to be delivered by electroporation or in formulation.
According to this paper, the dsRNA appears to be a double-stranded bond with the anti-sense RNA produced by using the product RNA as a template. Furthermore, this paper states that a full length anti-sense RNA can also be produced.
Those are statements reminiscent of the "Priongate" risks to me. In other words, this paper suggests that the RNA encoding "ORF 19" of the plasmid DNA of the Pfizer Omicron vaccine may be generated.
As mentioned in the below past tweet, ORF 19 includes 23 GxxxG-motifs and 144 G4-motifs. These are known as functional factors of the prion diseases.
We look forward to Kevin McKernan publishing his careful study of Endotoxin in various Jabs as he works to overcome interference in the LAL test caused by LNPs, pH, EDTA, HEPES, Phosphate Buffer and other contaminants arising from production.
Wow. Impressive work and very well explained.
What is going on with the magnesium levels? Looked at the EMA document and nothing specific was found. Bet the FDA knows all this. Plus it explains the EMA asking for further work on the DNAse 1.
Also, I have always wondered why BioNTech didn't use its new nifty cellulose based chromatography method which gets rid of 90% of the dsRNA? Maybe it wasn't enough or not enough RNA yield.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6444222/
Thank you.
The magnesium issue has not been considered yet, and I am currently at the stage of monitoring the verification since I'm currently busy with my day job and don't have enough time….
Thank you for presenting the paper. Katalin Karikó, BionTech, is one of the authors. I had a question in my mind about what dsRNA really is. There were some interesting points about the dsRNA production in this paper. That's the part below.
------
Phage RNA polymerases, including T7 RNA polymerase (T7RNAPol), transcribe the RNA with high fidelity from a DNA template containing the corresponding promoter. However, during the initiation of transcription, 5- to 11-nt-long RNA by-prod- ucts are generated as the enzyme aborts the synthesis with a certain probability.3,4 Considering that the T7RNAPol also has an obscure RNA-dependent as well as template-independent RNA polymerase activity,5–12 the short abortive RNA fragments and the 30 end of the full-length RNA can prime complementary RNA synthesis from the primary transcripts that leads to the generation of double-stranded RNA (dsRNA) contaminant. Recently, a promoter-independent transcription of full-length anti-sense RNA has been also reported as a novel mechanism of dsRNA generation in T7RNAPol-driven IVT reaction.13
The nuclei of human cells contain dsRNA that plays a role in nat- ural biological processes;14,15 however, when dsRNA enters the cells, from the extracellular milieu into their endosome or cytoplasm, it is sensed as a viral invader. All cells are capable of responding to dsRNA through sensors and effectors. Activation of dsRNA-depen- dent enzymes, such as oligoadenylate synthetase (OAS), RNA- specific adenosine deaminase (ADAR), and RNA-activated protein kinase (PKR), results in the inhibition of protein synthesis. In addi- tion, stimulation of the dsRNA sensors, e.g., Toll-like receptor 3 (TLR3), retinoic acid-inducible gene I (RIG-I), and melanoma dif- ferentiation-associated protein 5 (MDA5), leads to the secretion of different cytokines, including type I interferons, interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-a) (reviewed by Hart- mann16). Therefore, the elimination of dsRNA from the IVT mRNA is crucial to improve translation of the administered mRNA and to limit induction of cytokines, whether the mRNA products are intended for in vitro, ex vivo, or in vivo applications and to be delivered by electroporation or in formulation.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6444222/
------
According to this paper, the dsRNA appears to be a double-stranded bond with the anti-sense RNA produced by using the product RNA as a template. Furthermore, this paper states that a full length anti-sense RNA can also be produced.
Those are statements reminiscent of the "Priongate" risks to me. In other words, this paper suggests that the RNA encoding "ORF 19" of the plasmid DNA of the Pfizer Omicron vaccine may be generated.
As mentioned in the below past tweet, ORF 19 includes 23 GxxxG-motifs and 144 G4-motifs. These are known as functional factors of the prion diseases.
https://x.com/Patent_SUN/status/1699619448909713661?s=20
I would like to organize Priongate's substack someday.
Thank you.
Subtitle suggestion: Fun With Numerators and Denominators.
Thank you for sharing. Worth the read.
Excellent work and thanks very much for the mention.
Pfizer knew how to remove Double Stranded RNA Contaminants in 2011 but didn't do it for Covid19
https://geoffpain.substack.com/p/pfizer-knew-how-to-remove-double
We look forward to Kevin McKernan publishing his careful study of Endotoxin in various Jabs as he works to overcome interference in the LAL test caused by LNPs, pH, EDTA, HEPES, Phosphate Buffer and other contaminants arising from production.
https://geoffpain.substack.com/p/measuring-endotoxin-in-jabs-with
Thank you.