Genetic screening for single-cell variability modulators driving therapy resistance

Nat Genet. 2021 Jan;53(1):76-85. doi: 10.1038/s41588-020-00749-z. Epub 2021 Jan 4.


Cellular plasticity describes the ability of cells to transition from one set of phenotypes to another. In melanoma, transient fluctuations in the molecular state of tumor cells mark the formation of rare cells primed to survive BRAF inhibition and reprogram into a stably drug-resistant fate. However, the biological processes governing cellular priming remain unknown. We used CRISPR-Cas9 genetic screens to identify genes that affect cell fate decisions by altering cellular plasticity. We found that many factors can independently affect cellular priming and fate decisions. We discovered a new plasticity-based mode of increasing resistance to BRAF inhibition that pushes cells towards a more differentiated state. Manipulating cellular plasticity through inhibition of DOT1L before the addition of the BRAF inhibitor resulted in more therapy resistance than concurrent administration. Our results indicate that modulating cellular plasticity can alter cell fate decisions and may prove useful for treating drug resistance in other cancers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Cell Differentiation / genetics
  • Cell Line, Tumor
  • Cell Plasticity / genetics*
  • Cell Proliferation / genetics
  • Drug Resistance, Neoplasm / drug effects*
  • Drug Resistance, Neoplasm / genetics*
  • Genetic Testing*
  • Histone-Lysine N-Methyltransferase / genetics
  • Humans
  • Melanoma / drug therapy
  • Melanoma / genetics
  • Melanoma / pathology
  • Mice, Inbred NOD
  • Mice, SCID
  • Models, Biological
  • Molecular Targeted Therapy
  • Neoplasms / drug therapy
  • Neoplasms / genetics*
  • Neoplasms / pathology*
  • Proto-Oncogene Proteins B-raf / genetics
  • Transcription, Genetic


  • DOT1L protein, human
  • Histone-Lysine N-Methyltransferase
  • Proto-Oncogene Proteins B-raf