Researchers at ETH Zurich in Basel report they have used CRISPR-Cas technology to decipher how mutations in a cell’s genome affect its function. With their new approach, the researchers can generate thousands of cells with different gene variants in Petri dishes and determine which of the variants contribute to cancer development. The findings are published in in an article titled, “ ,” and led by ETH professor Randall Platt, PhD.
“Mutational scanning connects genetic variants to phenotype, enabling the interrogation of protein functions, interactions, and variant pathogenicity,” the researchers wrote. “However, current methodologies cannot efficiently engineer customizable sets of diverse genetic variants in endogenous loci across cellular contexts in high throughput. Here, we combine cytosine and adenine base editors and a prime editor to assess the pathogenicity of a broad spectrum of variants in the epithelial growth factor receptor gene (EGFR).
Using pooled base editing and prime editing guide RNA libraries, we install tens of thousands of variants spanning the full coding sequence of EGFR in multiple cell lines and assess the role of these variants in tumorigenesis and resistance to tyrosine kinase inhibitors.” Scientists already had the ability to make individual changes to the genome of cells. However, the ETH researchers modified one gene in two human cell lines in over 50,000 different ways, thereby creating a correspondingly large number of different .