DNA and RNA

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Tero
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Re: DNA and RNA

Post by Tero » Thu Apr 01, 2021 12:19 pm

RNA is not just any old molecule, and our RNA is protected inside the cell. The lipid package was the key to RNA survival before it enters a cell:
The impingement jet mixer, also known as the tea stirrer, works by simply pumping lipids in one side and mRNA in the other, forcing them together with 400 pounds of pressure. That's what creates the lipid nanoparticle which is essentially the vaccine.

These aren't just any lipids, Pfizer/BioNTech had to design the right combination of four different lipids that would not only protect the mRNA on the way to cells, but then release the mRNA once it gets there.

https://www.cnn.com/2021/03/31/health/p ... index.html
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Re: DNA and RNA

Post by Tero » Wed Jun 09, 2021 10:28 am

mRNA vaccines are made in plasmids. See 1:40 of the video



here, start from 13 min..there is not much on the RNA there

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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Re: DNA and RNA

Post by Tero » Wed Jun 09, 2021 10:32 am

Contract manufacturer...a lot of detail...3 min on has the plasmid bit



https://en.wikipedia.org/wiki/Plasmid
Plasmids can also be classified into incompatibility groups. A microbe can harbour different types of plasmids, but different plasmids can only exist in a single bacterial cell if they are compatible. If two plasmids are not compatible, one or the other will be rapidly lost from the cell. Different plasmids may therefore be assigned to different incompatibility groups depending on whether they can coexist together. Incompatible plasmids (belonging to the same incompatibility group) normally share the same replication or partition mechanisms and can thus not be kept together in a single cell.
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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Re: DNA and RNA

Post by Tero » Wed Jun 09, 2021 10:56 am

All enzymes and reaction components required for the GMP production of mRNA can be obtained from commercial suppliers as synthesized chemicals or bacterially expressed, animal component-free reagents, thereby avoiding safety concerns surrounding the adventitious agents that plague cell-culture-based vaccine manufacture. All the components, such as plasmid DNA, phage polymerases, capping enzymes and NTPs, are readily available as GMP-grade traceable components; however, some of these are currently available at only limited scale or high cost. As mRNA therapeutics move towards commercialization and the scale of production increases, more economical options may become accessible for GMP source materials.

GMP production of mRNA begins with DNA template production followed by enzymatic IVT and follows the same multistep protocol that is used for research scale synthesis, with added controls to ensure the safety and potency of the product16. Depending on the specific mRNA construct and chemistry, the protocol may be modified slightly from what is described here to accommodate modified nucleosides, capping strategies or template removal. To initiate the production process, template plasmid DNA produced in Escherichia coli is linearized using a restriction enzyme to allow synthesis of runoff transcripts with a poly(A) tract at the 3′ end. Next, the mRNA is synthesized from NTPs by a DNA-dependent RNA polymerase from bacteriophage (such as T7, SP6, or T3). The template DNA is then degraded by incubation with DNase. Finally, the mRNA is enzymatically or chemically capped to enable efficient translation in vivo. mRNA synthesis is highly productive, yielding in excess of 2 g l−1 of full-length mRNA in multi-gram scale reactions under optimized conditions.
https://www.nature.com/articles/nrd.2017.243
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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Re: DNA and RNA

Post by Tero » Mon Jun 14, 2021 12:21 pm

mRNA production in the first couple of minutes. This is a turning point. Much of new medicine will be from this technology.

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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Re: DNA and RNA

Post by Tero » Thu Jun 24, 2021 1:40 pm

Plasmids are round pieces of bacterial DNA. They are necessary for all mRNA production. Small companies making reagents for DNA or RNA will see quite a boom in the next few years.
A diversity of vaccine modalities are in development, in order to maximise the chance of developing at least one that can generate a safe and efficacious countermeasure to Covid-19. Two of these types, mRNA and DNA vaccines (nucleic acid vaccines), rely on plasmid DNA in their manufacture. Unlike traditional vaccines, which aim to elicit an immune response by introducing antigens via a weakened virus or protein, nucleic acid vaccines directly administer a part of the virus’ genetic code. These sequences are translated once in the body to produce viral proteins that elicit an immune response – most commonly for SARS-CoV-2, the “spike” protein.

Nucleic acid vaccines offer multiple advantages over traditional vaccines, the biggest of which, in the face of a pandemic, is the speed of production. Taking preclinical development as an example, traditional vaccines take on average a year and a half to reach Phase I [3]. In comparison, we have seen Covid-19 mRNA vaccines enter the clinic just a few months after the genetic sequence of SARS-CoV-2 was announced. This rapid pace of development makes it possible that the first vaccine approved for Covid-19 could also be the first nucleic acid vaccine ever approved for use in humans. Moreover, the speed of development allows nucleic acid vaccines a flexibility not possible with traditional vaccines, giving them distinct advantages in fighting novel viral pathogens as they emerge.

Many players are involved in the Covid-19 nucleic acid vaccine space, including big pharma, such as Pfizer, GSK and Sanofi, innovative biotechs, such as Biontech, Moderna, Curevac, Inovio and Touchlight, and academic groups, including Imperial College London. Current frontrunners are Moderna [4] and BioNTech/Pfizer [5], both of which are developing mRNA vaccines and have reached Phase III and Phase IIb/III respectively.

With clinical development well underway, attention is now firmly on scaling up manufacturing capacity to deliver the billions of doses that will be needed should these vaccines obtain regulatory approval and authorisation. Manufacturing mRNA and DNA vaccines requires significant quantities of DNA, which is used directly as the product in a DNA vaccine, or as a starting template for an enzymatic reaction for mRNA production. As an example, delivering one billion doses of an mRNA vaccine may require production of in excess of 1 kg of DNA. Using current plasmid DNA manufacturing techniques this is going to be a substantial feat and will require global collaborations between biotech, pharma and contract development and manufacturing organisations (CDMOs). To put it into perspective, Aldevron, the market leader in plasmid DNA manufacturing, recently noted that over half of the world's plasmid DNA manufacturing capacity will be required for just one Covid-19 mRNA vaccine [6].
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564888/
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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Tero
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Re: DNA and RNA

Post by Tero » Thu Jun 24, 2021 6:44 pm

From there, nearly all of the Pfizer DNA plasmid came from that Aldevron, from Fargo ND.
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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Re: DNA and RNA

Post by JimC » Thu Jun 24, 2021 9:22 pm

A Melbourne company, with state government assistance, is working towards producing a mRNA vaccine:

https://www.abc.net.au/news/2021-06-20/ ... /100229294
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Re: DNA and RNA

Post by Tero » Wed Jun 30, 2021 9:36 pm

Introducing RNA or DNA for other than vaccine use going to have complications. If you can inject it into an eyball, fine. To treat the eye.
https://blogs.sciencemag.org/pipeline/a ... maybe-isnt
https://esapolitics.blogspot.com
http://esabirdsne.blogspot.com/
Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Re: DNA and RNA

Post by Woodbutcher » Mon Jul 05, 2021 6:44 am

You eat about 150000 km of DNA in an average lunch.
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Re: DNA and RNA

Post by Svartalf » Mon Jul 05, 2021 9:17 am

who cares, it all gets turned to soup once you start chewing, and even more so once down in the stomach.
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Re: DNA and RNA

Post by JimC » Mon Jul 05, 2021 9:36 am

The atoms of beef DNA become atoms of JimC DNA.

A great honour, IMO... :tea:
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Re: DNA and RNA

Post by Sean Hayden » Sat Jul 10, 2021 6:48 pm

:lol:

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Tero
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Re: DNA and RNA

Post by Tero » Fri Jul 30, 2021 10:20 am

It's a year old, but pretty good review of vaccine:
https://internal-journal.frontiersin.or ... 85354/full
https://esapolitics.blogspot.com
http://esabirdsne.blogspot.com/
Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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Tero
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Re: DNA and RNA

Post by Tero » Fri Jul 30, 2021 7:23 pm

Genes code for peptides. One of the most studied peptides is insulin. If it is going to act like insulin in mammals, it has to have mostly the same sequence. However, it has departed from that in some species.
Insulin sequences from the guinea pig (Cavia porcellus) and relatives (rodents of the suborder Hystricomorpha) are well known for having insulin sequences with highly divergent sequences (39, 40). The biological activities of these insulins also differ, acting more as a growth factor than as a metabolic hormone (41, 42). These changes, in sequence and function, have been accompanied with an acceleration of the rate of evolution of the insulin protein sequence in the guinea pig and relatives (40, 43, 44). Similar, but less dramatic, episodes of accelerated evolution of insulin sequences have been observed in some species of New World monkeys (44, 45), species that have insulin hormones with lower potency (46).
https://www.frontiersin.org/articles/10 ... 49255/full
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Said Peter...what you're requesting just isn't my bag
Said Daemon, who's sorry too, but y'see we didn't have no choice
And our hands they are many and we'd be of one voice
We've come all the way from Wigan to get up and state
Our case for survival before it's too late

Turn stone to bread, said Daemon Duncetan
Turn stone to bread right away...

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