In epoca vaccinazioni Covid-19 i genitori si sono fidati delle vaccinazioni a mRNA ed oltre a vaccinarsi loro stessi hanno fatto vaccinare i propri figli con un conseguente aumento delle morti preadolescenziali e adolescenziali oltreché prenatali e neonatali. I dati ci sono e sono sotto gli occhi di tutti...

Efficacia e sicurezza dei più recenti vaccini a mRNA sono state pesantemente smentite dall'enorme numero di morti e di persone invalidate dai vaccini Pfizer, Moderna, Astrazeneca ed altri. L'insorgenza di miocarditi e di pericarditi o di entrambe in vario grado è evidentissima; il numero di patologie autoimmuni e di tumori fulminanti insorti in soggetti in età infantile, adolescenziale, giovanile adulta è in notevole crescita in tutti i Paesi in cui la popolazione ha aderito massicciamente alle vaccinazioni a mRNA. Autoimmunity Reactions

Review coordinated by Life Science Editors Foundation

Reviewed by: Dr. Angela Andersen

Potential Conflicts of Interest: None

Background: * mRNAs in polarized cells often have a distinct spatial localization patterns that enable localized protein production * In non-polarized cells, mRNAs encoding membrane and secretory proteins are predominantly translated on the endoplasmic reticulum (ER), some mRNAs are enriched on the mitochondrial surface, some mRNAs are bound to the RNA-binding protein (RBP) TIS11B at the surface of the rough ER in "TIS granules". * The translation of specific mRNAs in TIS granules allows assembly of protein complexes that cannot be established when the mRNAs are translated on the ER but outside of TIS granules (physiological relevance). * The canonical rough ER (CRER) is distinct from the TIS granule ER (TGER), and both are distinct from the cytosol.

Questions: * Do mRNAs that encode non-membrane proteins differentially localize to the ER or the cytosol? (in steady state) * Does the amount of protein synthesis differ depending on the subcytoplasmic location of an mRNA?

Summary: * A third of mRNAs that encode non-membrane proteins have a biased localization to TGER or CRER, indicating that the ER membrane is a general site of translation for both membrane and non-membrane proteins. * 52% of mRNAs that encode non-membrane proteins have a biased mRNA transcript localization pattern towards a single cytoplasmic compartment. the TGER, CRER or cytosol. * The localization at the TGER or CRER was largely controlled by a combinatorial code of AU-RBPs at the 3'UTR. TIS11B promotes mRNA localization to TGER and TIA1/L1 to CRER. * LARP4B bound to the 3'UTR promotes cytosolic localization. * The location of translation has an independent effect on protein levels independent of the RBPs/3'UTR: redirecting cytosolic mRNAs to the rough ER membrane increased their steady-state protein levels by two-fold, indicating that the ER environment promotes protein expression. * Compartment-enriched mRNAs differed in their mRNA production and degradation rates, as well as functional classes and levels of their encoded proteins. Therefore the cytoplasm is partitioned into different functional and regulatory compartments that are not enclosed by membranes. * low-abundance proteins are translated in the TGER region. mRNAs encoding zinc finger proteins and transcription factors were substantially enriched at the TGER. These gene classes are usually expressed at lower levels than others.. This localization may regulate protein complex assembly (membrane proteins that are translated in the TGER domain establish protein complexes that cannot be formed when the proteins are translated on the CRER). The TGER may ensure that low-abundance mRNAs are effectively translated into low-abundance proteins. * mRNAs that are the most stable and encode the most highly expressed proteins are enriched on the CRER and include helicases, cytoskeleton-bound proteins, and chromatin regulators, overturning the idea that most non-membrane protein-encoding mRNAs are translated in the cytosol. * mRNAs overrepresented in the cytosol had the highest production and degradation rates and were enriched in proteins involved in mRNA processing and translation factors, whose abundance levels require tight control.

Advance: Evidence for functional compartmentalization of non-membrane mRNA protein expression in the cytosol vs ER. In steady state, general localization of mRNAs to the ER promotes high protein levels.

Significance: Engineered 3'UTR sequences could potentially boost protein expression by localizing mRNAs to the ER in experimental settings, for vaccines etc.

Remaining questions/points: * How does the rough ER stimulate protein expression? * Does the mRNA localization affect complex formation and/or function of non-membrane proteins? * Does this occur in cells other than HEK293T? * Is this regulated?

mRNA needs to be assisted across the NPC. Like protein, also classed as facilitated diffusion. * mRNP exporter combines with mRNA with poly A tail, by interacting with FG repeats * mRNA moves through NPC * Dpb5 is an RNA helicase * Straightens the mRNA secondary structure and allows passage, removes proteins on the strand (NXT1, NXF1) * mRNP exporter proteins dissociate from the mRNA. * mRNA is now in cytoplasm

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These 2 articles form an informational island on covid19 mRNA vaccines.

Diagrams of the presence of proteins versus time in various administration methods in mouse

Where does all the mRNA end up, according to this 2015 study on mice](https://www.ncbi.nlm.nih.gov/labs/pmc/articles/PMC4624045/)?

Here we learn stuff not reorted in the paper.

Variants that show reduced sensitivity to NAbs don't necessarily mean mRNA vaccine failure

New variants may emerge that show reduced sensitivity to NAbs.

This may not result in vaccine failure because:

1. The mRNA vaccines induce such a strong NAb response, there will be enough spare capacity to deal with the virus.
2. The mRNA vaccines also induce other virus specific protection such as helper T cells and cytotoxic T cells, which may not be affected by the reduction in NAb sensitivity.

Most vaccines for respiratory illnesses are leaky.

The efficacy the mRNA vaccines showed in preventing asymptomatic transmission was therefore a welcome surprise.

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Vaccine's halg-life was measured just below a day for mice muscles.

Spleen & liver had x30 less mRNA concentration than Lymph nodes closest to the vaccine site, and x70 less the injected muscle.

Lymph-node mRNA half-life is a day.

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Safety of mRNA Liquid Nanoparticles

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This is the process of a complementary mRNA copy of a single gene on the DNA that is created in the nucleus. The mRNA is smaller than the DNA so it can carry the genetic code into the ribosome and into the cytoplasm that enables the protein creation.

This is an important mechanism that cells can respond to their environment and extracellular cues. Cells sense their environment and modify genes, mRNA splicing, protein expression and protein modifying to respond to these cues.

CohenNov. 16, J., 2020, & Am, 7:00. (2020, November 16). ‘Just beautiful’: Another COVID-19 vaccine, from newcomer Moderna, succeeds in large-scale trial. Science | AAAS. https://www.sciencemag.org/news/2020/11/just-beautiful-another-covid-19-vaccine-newcomer-moderna-succeeds-large-scale-trial

This is a single strand on an RNA molecule that leaves the the nucleus of a cell in order to relocate to the cytoplasm. This is where the mRNA can help create the protein for the cell in a process known as protein synthesis. The mRNA takes in information passed into it by DNA and decode it for the ribosomes to make more protein for the cell to live on.

Corbett, K. S., Edwards, D. K., Leist, S. R., Abiona, O. M., Boyoglu-Barnum, S., Gillespie, R. A., Himansu, S., Schäfer, A., Ziwawo, C. T., DiPiazza, A. T., Dinnon, K. H., Elbashir, S. M., Shaw, C. A., Woods, A., Fritch, E. J., Martinez, D. R., Bock, K. W., Minai, M., Nagata, B. M., … Graham, B. S. (2020). SARS-CoV-2 mRNA vaccine design enabled by prototype pathogen preparedness. Nature, 1–8. https://doi.org/10.1038/s41586-020-2622-0