New “anti-CRISPR” discovered in viruses...It could be an “off” switch for gene editing and create better weapons against superbugs.

Anewly discovered “anti-CRISPR” system in viruses could help us control CRISPR gene-editing technologies and lead to better weapons against antibiotic-resistant superbugs.

The background: Your immune system has ways to remember viruses you’ve encountered before and help defend you if they appear again — and, amazingly, some single-celled bacteria do, too.

These microbes evolved the CRISPR system to protect themselves from the viruses that infect bacteria, called phages. The bacteria will take a little bit of a virus’ DNA sequence and add it to their own genome, like a memory of past infections. They then create CRISPR-associated (Cas) proteins containing copies of the viral genetic sequence, written in RNA.

Those Cas proteins then patrol the bacteria, waiting for a virus with the matching DNA sequence to show up again. If it does, the CRISPR system slices through the virus’ DNA to disable it.

The challenge: That’s how CRISPR works in nature, but researchers have learned how to program the systems to match up with any desired genetic sequence — they can then deploy it to make precise edits to all kinds of DNA to treat diseases, improve crops, and more.

To take full advantage of CRISPR as a gene-editing tool, though, we need to figure out better ways to control it. Ideally, we’d want to be able to deploy the system, and then turn it off once it’s edited enough cells (or if it starts making edits in places it’s not supposed to).

A new anti-CRISPR: Researchers from the University of Otago and the University of Copenhagen have now discovered a new “anti-CRISPR” system that phages have evolved to get around bacteria’s defenses.

“It’s like an evolutionary arms race,” said co-first author David Mayo-Muñoz. “Bacteria have CRISPR-Cas, so the phages have developed anti-CRISPRs, which enables them to block the immune complexes of the bacteria.”

Researchers have found ways to use previously discovered anti-CRISPR systems as off-switches for CRISPR gene-editing systems, but those anti-CRISPRs all used proteins to prevent a bacteria’s CRISPR system from working.

This is the first anti-CRISPR system they’ve seen where the virus creates RNA that mimics the RNA in its host’s CRISPR system. The viral RNA then binds directly to the bacteria’s Cas proteins to disable them.

“This molecular mimic ruins the defenses of the bacteria and the function of the system; it’s basically a decoy,” said co-lead researcher Peter Fineran.

Looking ahead: The researchers believe it will be easier to find and use new RNA-based anti-CRISPRs than protein-based ones, and the more we can study, the better equipped we’ll be to use the systems to develop off-switches for CRISPR gene-editing tech.

Research into these anti-CRISPRs could also help us create new phage therapies, which use viruses that aren’t harmful to people to kill infections caused by antibiotic-resistant bacteria.

“If we’re able to equip phages with anti-CRISPR strategies, they would be more effective at taking over bacterial populations and killing nasty bugs,” researcher Rafael Pinilla-Redondo told Nature Podcast.

CRISPR has nothing to do with the mRNA technology used to create the Covid vaccines.

The plasmids that encode the modified spike protein mRNA were created using traditional gene engineering techniques that date back to the 1970s or earlier.

CRISPR has been touted as a game-changer when, in reality, it’s just another tool in our genetic-engineering toolbox.

Of course the Covid-19 vaccines are vaccines. The purpose of a vaccine is to show your immune system what a specific foreign protein (i.e. antigen) looks like so that the immune system can develop an antigen-specific T-cell, B-cell, and antibody response. Those T-cells, B-cells, and antibodies then circulate throughout your body looking for that antigen. When they see it, they immediately move to disable and destroy it.

In the specific case of Covid-19, the immune response induced by the vaccine specifically targets the spike protein. So when you are exposed to Covid viruses that are covered with spike protein, those T-cells, B-cells, and antibodies immediately go to work. If your immune response is good enough, they’ll destroy all of the invading virus particles without you ever knowing you were exposed. With a weaker immune response, you might still get sick but you have a head start in combating the disease before it can damage you. Your chance of long-term health impairments or death from Covid is significantly reduced if you have pre-trained your immune system with a vaccine.

“In summary, our work establishes mRNA-mediated delivery of CRISPR/Cas9-based tools”

What that means is that they transfected cells with mRNA encoding the Cas9 gene. This is similar to the process following injection of the mRNA based vaccines. The mRNA enters the cell, causes the cell to make the protein encoded in the mRNA for a while, after which the cell destroys the mRNA. mRNA has a half-life of hours.

The fact that inserting of mRNA encoding Cas9 and mRNA encoding a modified SARS-CoV-2 spike protein basically use the same method does not mean that CRISPR was in any way involved in creating the Covid-19 vaccines.

Also, according to the Cell paper that you linked, the Cas9 enzyme functions because another RNA (not an mRNA) was transfected into the cells along with the Cas9 mRNA. That other RNA contained a mutation matching the specific site in the DNA that the researchers wanted to modify. The only mRNA present in Covid vaccines does not encode Cas9 and there is no RNA to cause a point mutation in the DNA.

“The Pfizer and Moderna COVID vaccines are the first vaccines to be activated by mRNA. These vaccines build on the breakthroughs of the gene-editing technology known as CRISPR.”

The NPR interview does not tie CRISPR to the creation of the mRNA vaccine at all. It discusses the vaccine in the context that some of the same techniques are used in delivering mRNA into cells. As far as the mRNA vaccine development being dependent on breakthroughs from CRISPR development, I have my doubts. The technologies were developed more or less in parallel. The fact that some of the methods are similar (for example, delivering mRNA into cells) does not mean that vaccine development was dependent on CRISPR development.

Mostly, that interview appears to be more CRISPR hype. Often, scientists who have discovered a new method of doing something will hype it in order to increase stockholder interest in their biotech company and increase sales of kits they developed to use the method. Is CRISPR really worth all the hype? I doubt it. As I said, it is just another tool in a huge molecular biology toolbox. Everything that can be done with CRISPR was possible using other enzymes and methods prior to the discovery of CRISPR.

Okay.

At best mRNA from anticovid injection is based on manipulating host mRNA traffic from the gene to protein synthesis.

Your first paragraph sort of summarizes the vaccine action. However, no host cell mRNA is involved in the production of modified spike protein. This is because the human genome does not contain any spike protein genes. The mRNA from the vaccine uses the cell's protein synthesis machinery in order to produce the modified spike protein. Within a few hours, the foreign mRNA is destroyed in the same manner as the thousands of host mRNA molecules are destroyed. Once the foreign mRNA is destroyed, the cell makes no more spike protein.

The interaction between the transposon, and retrotransposon system are overlooked.

This is primarily a concern during cell division occurring during a viral infection. The retrotransposon system becomes active only during cell division. If the cell happens to be infected with an RNA virus (like coronavirus) at the time, it is theoretically possible for the retrotransposon enzymes to make DNA using the RNA as a template and then to insert it into the genome. Theoretically possible means that I do not know that this has actually been demonstrated to occur during a coronavirus infection; it has only been demonstrated using cells in a lab.

The degradation of the mRNA could possibly yield microRNA or picoRNA snippets which can be active controllers of DNA activity.

Nope. As I already pointed out, the human genome does not contain a SARS-CoV-2 spike gene. Without such a gene, there would be no place on the genome for fragments of the mRNA to attach to. Furthermore, RNA destroying enzymes love nothing more than to chop up RNA into its component nucleotides. One of the biggest challenges for scientists working with RNA is to prevent its destruction. RNA is notoriously unstable.

Why any ethical drug developer would think it is good idea to try manipulation ofgenetic machinery in order to create an antigenic, host derived spike protein is beyond me.

No drug developer did anything to manipulate the human genome. All they did was to take advantage of the cell's natural ability to make protein using mRNA templates.

I should note that during a virus infection, the virus forces your cells to make virus mRNA and virus protein in order to form new virus particles; once the cell is full of virus particles, it bursts open and releases them to neighboring cells and into the blood where the new virus particles can travel to distant parts of the body and infect more cells. What this means is that there is nothing new about human cells using virus mRNA; it happens every time you have a virus infection.

The difference with the vaccine mRNA is that it only contains mRNA coding for the modified spike protein but none of the mRNA encoding the other 28 virus proteins. Thus, it is not possible for cells to make viruses following a vaccine injection. This means that cell death is not an inevitable outcome after vaccination, like it is after virus infection.

All they had to do was kill the virus and use that material as an “old school” vaccine. Or synthesis nucleocapsid and spike protein using recombinant technology in vitro.

The reason mRNA vaccine technology was developed is because all of those "old school" vaccine technologies are very time and resource consuming. Growing and killing a virus to make a vaccine is time-consuming (and somewhat dangerous, since it involves work with an active pathogen). Any time the circulating virus strain changes, it becomes necessary to isolate and grow the new virus and then develop and test a new vaccine made with killed virus. This takes time. This is how influenza vaccines are updated, and it takes months between identifying the need to develop a new influenza vaccine and being able to actually deliver the vaccine to patients.

The synthesis of virus antigen in the lab can be done, but it takes more steps and reagents than just making mRNA. It still requires a plasmid encoding the mRNA, but it additionally requires all of the protein production machinery contained in cells. I used to use extracts of rabbit reticulocyte cells to make proteins in the lab. Those extracts were expensive, not to mention the fact that rabbits had to die so that I could get those extracts. And they weren't just killed humanely--they were poisoned to make them sick days before they were killed. This was done to cause the rabbits to make an abundance of reticulocytes. How many rabbits would have to be poisoned and killed in order to synthetically produce enough modified spike protein to vaccinate hundreds of millions of people? Are there even that many pathogen-free research rabbits in existence?

The thing about the mRNA technology is that making the vaccine only requires the creation of a plasmid--which is a fairly rapid and straightforward process--and then growing that plasmid in bacteria so that they will make mRNA. Any time the circulating virus strain changes, it is a straightforward process to make a new plasmid encoding a new modified spike protein. Bacteria can be grown in huge vats and they grow very fast. They are fed a milk protein. They produce a lot of mRNA which is then cleaned and formulated into vaccines. It is probably the most rapid and straightforward vaccine technology developed so far.

No dear, you denied that there was any connection between mRNA and CRISPR when it is an established fact that there is. It is also an established or stipulated fact that the COVID shot implements CRISPR technology.

Please, please, please stop. You only demonstrate just how little you comprehend molecular biology.

I'm reminded of the time I tried to tutor a woman who had absolutely no grasp of simple mathematics. She was, quite literally, impossible to teach. She could not understand even the simplest of mathematical problems.

I will explain (again), but I don't think you can understand.

mRNA is the intermediary between DNA and protein. Every single cell in every living organism constantly makes and destroys mRNA. The purpose of the mRNA is to carry the genetic information from the DNA to the protein-making machinery, which is called a ribosome. The ribosome "reads" the mRNA in order to make a protein. The modified SARS-CoV-2 spike protein is a protein. The Cas9 enzyme used in CRISPR is a protein. Since they are both proteins, they are both made by ribosomes reading mRNA. The fact that they (and millions of other proteins) are all made by the same cellular systems does NOT mean they are connected.

Once again, the Cas9/CRISPR system has absolutely nothing to do with the Covid vaccine. The Covid-19 vaccine does not contain Cas9 mRNA or the supplemental RNAs that Cas9 uses in CRISPR mediated DNA modification. And the Cas9/CRISPR system contains no spike protein mRNA.

So everyone who says the Covid shot uses mRNA and CRISPR lied? Pfizer, Abbott, J&J, NPR, CDC, FDA...they’re all wrong and you’re right?

Literally NO ONE has said that the Covid-19 mRNA based vaccines use CRISPR.

The only one who keeps insisting that CRISPR tools are in the vaccine is YOU. I do not think you are intentionally lying, since you actually have to know the truth before you can lie.

I do think that you read some antivax screed somewhere that probably talked about CRISPR in such a way as to imply that the gene-editing tool CRISPR is contained in vaccines, without actually saying that it is.

You ate breakfast this morning, didn't you? Your breakfast was chock-full of mRNA. OMG! THIS MEANS YOUR BREAKFAST IS FULL OF CRISPR AND MODIFYING YOUR GENES! OMG! PANIC! PANIC!

Now, will you please just STOP. Your absolute ignorance and inability to understand science is embarrassing to witness. Profoundly. I don't even know you and I'm cringing with embarrassment for you.

Disclaimer: Opinions posted on Free Republic are those of the individual posters and do not necessarily represent the opinion of Free Republic or its management. All materials posted herein are protected by copyright law and the exemption for fair use of copyrighted works.