U.S. researchers have introduced a significant upgrade of the CRISPR gene processing system, which, unlike its predecessor, can target high-precision specific genes and make completely reversible changes.
The new gene editing technology, developed by Whitehead Institute member Jonathan Weissman and the University of California San Francisco assistant professor Luke Gilbert, is called CRISPRoff.
It is capable of controlling a gene with laser-like precision, while leaving the broader piece of DNA unchanged. Furthermore, the modifications are not only stable enough to be inherited by hundreds of cell divisions, but are also completely reversible.
‘We can do this for multiple genes at the same time without any DNA damage, with a great deal of homogeneity and in a way that can be reversed. It is an excellent tool for controlling gene expression, ”says Weissman.
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The significant upgrade of the CRISPR-Cas9 technology is partly funded by the Defense Advanced Research Projects Agency (DARPA) and has been in development for over four years.
“CRISPRoff ultimately works as envisioned in a science fiction way,” says fellow senior author Gilbert. “It’s exciting to see it work so well in practice.”
The original CRISPR technology uses the DNA cleavage protein called Cas9, which can target specific genes in human cells and snip the DNA strand, which then recovers once the required gene has been removed. However, this process is permanent and depends on the cell’s own repair mechanisms to clean things up afterwards, which can lead to bad, unwanted, additional outcomes.
The researchers saw an opportunity in which genes could be silenced or activated due to chemical changes to the DNA strand, without any drastic and permanent changes.
The new CRISPRoff technology enables scientists to write a genetic program that can be remembered and replicated indefinitely by the cell, and the secretion or activation of different genes, through numerous cell divisions, in a method that they say is safer and is more effective than the original CRISPR.
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Gene silencing is usually based on a process known as methylation and works by adding chemical labels to certain parts of the DNA strand. The CRISPRoff researchers have created a small protein machine that can stick methyl groups to the required areas of the DNA strand, thus ‘silencing’.
When they want to reactivate the specific genes, they administer enzymes that remove these methyl groups, according to a method they naturally call CRISPRon.
In tests on pluripotent stem cells, which can turn into any type of cell, the researchers succeeded in almost completely silencing the expression of the Tau protein, which is strongly involved in the onset of Alzheimer’s disease.
“What we have shown is that it is a viable strategy to silence Tau and prevent proteins from being expressed,” says Weissman, adding that the way to give it to an adult has not yet been fully identified. should be, but it shows early promise. .
They have compared the delivery potential of CRISPRoff in humans with the newly developed RNA technology that forms the basis of both the Moderna and BioNTech coronavirus vaccines, but their gene editing technique will have to undergo much more careful before it can even begin. . clinical trials in human hosts.
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