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The introduction in 2007 of the gene editing tool, called CRISPR, is a revolution in the field of medical science and cell biology. However, even if the outlook is good, the launch of CRISPR was followed by a debate, focused in particular on the ethical issues and on the degree of precision and side effects of the technology.
However, in a new study published in the scientific journal Cell Researchers at the Protein Research Center of the Novo Nordisk Foundation described the operation of a CRISPR technology, called Cas12a, up to the molecular level. This helps refine the process of editing the genes to obtain only the desired effects.
"If we compare CRISPR to a car engine, we created a complete 3D engine map, which allowed us to understand how it works. This knowledge will allow us to refine the CRISPR engine and make it work in different ways – as a Formula 1 racer and off-road truck, "says Professor Guillermo Montoya of the Novo Nordisk Foundation Protein Research Center .
Molecular film
The researchers used a cryo-electron microscope to map the technology. The cryoEM facilities recently inaugurated at the University of Copenhagen have developed state-of-the-art technology allowing researchers to take photographs of the different forms of the molecule when CRISPR-Cas12a cuts the strand of DNA.
They have combined it with a fluorescence microscopy technique called "single molecule FRET" that directly observes the motions of the molecules and the sequence of events for each individual protein.
Among other things, this sequence of events revealed to researchers that three "elements" of the CRISPR tools needed to change shape in order for the DNA to be properly cut.
"Our new study shows the precise series of events in the genome leading to gene editing. These three "parts" that change work as security checks at airports. To proceed, you must perform all the checks and in the right order, "says Associate Professor Nikos Hatzakis of the Department of Chemistry and Center for Nanoscience.
More editing than desired
According to the researchers, their new findings may explain why CRISPR technology can have side effects on the genome. Once the DNA strand cut, the three "security checks" remain open.
The process may therefore take longer than expected because the mechanisms behind gene editing continue to work and may cause genetic changes.
However, researchers are now waiting for their new knowledge to put an end to this situation. They believe this technology can be used to immediately refine gene modification technology.
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The study is funded by the Novo Nordisk Foundation, the Carlsberg Foundation and the Villum Foundation.
Both research groups are members of the Integrative Structural Biology Cluster (ISBUC) of the University of Copenhagen.
Read the study: "The conformational activation promotes CRISPR-Cas12a catalysis and resetting endonuclease activity" to the address Cell.
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