The identification of functional regions of MEK1 using CRISPR tiling screens

Commun Biol. 2025 Apr 24;8(1):656. doi: 10.1038/s42003-025-07966-4.

Abstract

CRISPR tiling screen is a powerful tool to identify protein regions relevant to its biological function. Understanding the functional relevance of the regions of target protein is of great help for structure-based drug discovery. Studying the drug resistance mechanisms of small-molecule inhibitors is important for the development and clinical application of the compounds. Using MEK1 and MEK inhibitors as example here, we demonstrate the utility of CRISPR tiling to identify regions essential for cancer cell viability and regions where mutations are resistant to MEK inhibitors. We study the drug resistance mechanisms of the regions and discussed the potential, as well as limitations, of applying the technology to drug development. Our findings demonstrate the value and prompt the utilization of CRISPR tiling technology in structure-based drug discovery.

MeSH terms

  • CRISPR-Cas Systems*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Drug Discovery / methods
  • Drug Resistance, Neoplasm / genetics
  • Humans
  • MAP Kinase Kinase 1* / antagonists & inhibitors
  • MAP Kinase Kinase 1* / chemistry
  • MAP Kinase Kinase 1* / genetics
  • MAP Kinase Kinase 1* / metabolism
  • Mutation
  • Protein Kinase Inhibitors / pharmacology

Substances

  • MAP Kinase Kinase 1
  • MAP2K1 protein, human
  • Protein Kinase Inhibitors