Molecular Cell: New gene editing technology allows CRISPR-Cas9 to edit DNA sequentially

New gene editing technology allows CRISPR-Cas9 to edit DNA sequentially.  In a new study, researchers from the University of Illinois at Chicago have discovered a new gene editing technology that can edit sequences over time Cut or edit for programming. Related research results were recently published in the journal Molecular Cell, with the title of the paper “Sequential Activation of Guide RNAs to Enable Successive CRISPR-Cas9 Activities”.

CRISPR is a gene editing tool that allows scientists to change the DNA sequence in cells and sometimes add desired sequences or genes. CRISPR uses an enzyme called Cas9, which acts like scissors, cutting precisely at the desired position in the DNA. Once cut, the way the cell repairs DNA breaks will be affected, resulting in different changes or editing of the DNA sequence.

The discovery of the gene editing function of the CRISPR system was described in the early 2010s. In just a few years, scientists have become fascinated by such a technique: in experiments, guiding CRISPR to target almost all DNA sequences in the cell, or to target many different sites in the cell.

Bradley Merrill, co-corresponding author of the paper and associate professor of biochemistry and molecular genetics at the University of Illinois at Chicago School of Medicine, said, “One disadvantage of the currently available CRISPR-based editing systems is that all editing or cutting is done at once. There is no way. Guide them so that they happen sequentially one after another.”

The new method developed by Merrill and colleagues involves the use of special molecules called guide RNAs (gRNAs) that guide the Cas9 enzyme inside the cell and determine the precise DNA sequence where Cas9 will cut. They call their specially designed gRNA molecules “proGuide”, which allow the use of Cas9 to program and edit DNA.

Although proGuide is still in the prototype stage, Merrill and his colleagues plan to further develop their concept and hope that scientists can use the technology soon.

Merrill said, “The ability to pre-program Cas9 sequential activation at multiple sites introduces a new tool for biological research and genetic engineering. The time factor is a key factor in human development and a key factor in disease progression, but currently The method of genetic research on these processes does not make effective use of the time factor. Our system allows gene editing in a pre-programmed manner, allowing scientists to better study time-sensitive processes, such as how cancer develops from several genes. What develops in the mutation and how the order in which these mutations occur affects this disease.”

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