New CRISPR Method Allows Scientists to Understand Impact of Subtle Mutations – Columbia University Irving Medical Center

A new method that cleverly uses CRISPR to simultaneously make thousands of point mutations in human genes is helping researchers at Columbia University Vagelos College of Physicians and Surgeons understand how subtle changes to genes contribute to cancer and other diseases.

The new research is featured in "CRISPR base editor screens identify variant function at scale,"a perspective published in Molecular Cell, and in "Finding function with base editing screens," in Nature Reviews Genetics.

The method, created by a team led byAlberto Ciccia, PhD, associate professor of genetics and development in the Herbert Irving Comprehensive Cancer Center, was designed to study genes involved in DNA repairbutcould be applied to the study of any type of gene.

The findings,published in the Feb. 18 issue of Cell, could lead to precision medicines tailored to a patients exact genetic abnormality.

Researchers trying to understand how a gene impacts disease have generally been limited to making changes that disable an entire gene and its respective protein. Although this can reveal the genes dominant function, it doesnt tell us about all the other functions it may have, Ciccia says.

With a new type of gene editingcalled CRISPR-dependent cytosine base editinga single cytosine (C) in a gene can be replaced by a single thymine (T).

With base editors, we can make subtle changes in any part of the gene, much like what happens in nature, Ciccia says.

Making the technique even more powerful, the Ciccia laboratory has devised a way to make cytosine base edits across an entire gene and in dozens of genes at the same timea high-throughput approach that will greatly accelerate the discovery and analysis of clinically important mutations.

Using the new screening method, Ciccia and his team made thousands of mutations in 86 different DNA repair genes and exposed cells with the mutations to DNA-damaging agents to determine if the point mutations altered the function of these genes.

DNA repair genessense and correct errors in DNA, but mutations within the repair genes can hinder this natural proofreading mechanism. Genetic alterations of DNA repair genes have been implicated in breast and other cancers, along with other human genetic disorders.

The screen successfully identified more than 100 new, likely pathogenic mutations in DNA repair genes, including several in the breast cancer genes, BRCA1 and BRCA2.The full results of the screen are available on the Ciccia laboratorys website.

We anticipate this resource will expedite studies of DNA repair genes in human disease, Ciccia says, and that base-editing screening technologies will help researchers determine the causes of many different human diseases and accelerate the path to precision medicine.

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New CRISPR Method Allows Scientists to Understand Impact of Subtle Mutations - Columbia University Irving Medical Center