CRISPR prime editing for unconstrained correction of oncogenic KRAS variants

Abstract

KRAS is the most commonly mutated RAS family gene and is a primary cause of the occurrence of several types of cancer. However, KRAS mutations have several unique and diverse molecular identities, making it difficult to find specific treatments. Here, we developed universal pegRNAs which can correct all types of G12 and G13 oncogenic KRAS mutations with CRISPR-mediated prime editors (PEs). The universal pegRNA successfully corrected 12 types of KRAS mutations, accounting for 94% of all known KRAS mutations, by up to 54.8% correction frequency in HEK293T/17 cells. We also applied the universal pegRNA to correct endogenous KRAS mutations in human cancer cells and found that G13D KRAS mutation was successfully corrected to wild-type KRAS sequences with up to 40.6% correction frequency without indel mutations. We propose prime editing with the universal pegRNA as a ‘one–to–many’ potential therapeutic strategy for KRAS oncogene variants.

A CRISPR prime editing approach corrects all types of G12 and G13 oncogenic KRAS mutations using a universal pegRNA, demonstrating the potential of applying this system to KRAS gene therapy through a oneto-many approach.