Researchers understand the processes that affect the efficiency of the CRISPR method

When it comes to the ability of genetic engineering, the only restriction is the human imagination, particularly since the discovery of CRISPR technology.

A new Danish research project will help to fine-tune the technique and pave the way for more sophisticated and accurate use of “genetic scissors.”

Our study shows that, by better understanding the CRISPR/Cas9 protein and its gRNA component, we can more accurately hit and cut the DNA and thereby optimize the effectiveness of gene modification.”

Yonglun Luo, Associate Professor, Department of Biomedicine, Aarhus University

Revolutionary technology

Scientists found a protein in bacteria that can cut DNA and uses a so-called guide RNA to determine where the DNA needs to be cut 10 years ago. Since then, the CRISPR technique has been heralded as a rebellion in gene technology.

CRISPR allows the removal or adding of specific genes in any living organism, from bacteria to plants to humans.

CRISPR technology also makes it feasible to cure diseases by precisely correcting errors in human genes.

This means that the innovation has nearly limitless applications in basic research, public health, agriculture, and medicine.

Preventing unintended modifications

The technology’s implementation will necessitate that the technique is useful and accurate so that researchers only reach the desired, rather than unintended, gene modifications.

Scientists from the University of Copenhagen and Aarhus University used an energy-based model to determine the processes governing CRISPR-Cas9 activity and specification to truly comprehend the processes that affect the efficiency of the CRISPR method.

This model allows researchers to create gRNA components that improve the method’s effectiveness while minimizing unintended consequences, also known as “off-target effects.”

Unintended off-targets are a major concern when using the CRISPR method to treat diseases, and most of the tools for measuring off-targets have serious limitations and do not include the factors that we have discovered in our study. These discoveries have given us the key to designing CRISPR-gRNA with high effectiveness and precision.”

Yonglun Luo, Associate Professor, Department of Biomedicine, Aarhus University

Refining the method

To enhance the method’s potency and accuracy, the scientists behind the research will continue to evaluate the technique and the design of the gRNA component.

We’ll also try to find new methods of measuring on-target and off-target areas and developing innovative methods to address the off-target challenges that still limit our ability to use the CRISPR-Cas9 method.”

Yonglun Luo, Associate Professor, Department of Biomedicine, Aarhus University

Source:
Journal reference:

Corsi, G. I., et al. (2022) CRISPR/Cas9 gRNA activity depends on free energy changes and the target PAM context. Nature. doi.org/10.1038/s41467-022-30515-0.

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