Isolating live cell clones from barcoded populations using CRISPRa-inducible reporters

Nat Biotechnol. 2021 Feb;39(2):174-178. doi: 10.1038/s41587-020-0614-0. Epub 2020 Jul 27.

Abstract

We developed a functional lineage tracing tool termed CaTCH (CRISPRa tracing of clones in heterogeneous cell populations). CaTCH combines precise clonal tracing of millions of cells with the ability to retrospectively isolate founding clones alive before and during selection, allowing functional experiments. Using CaTCH, we captured rare clones representing as little as 0.001% of a population and investigated the emergence of resistance to targeted melanoma therapy in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Cell Line
  • Cell Separation*
  • Clone Cells / metabolism*
  • Female
  • Genes, Reporter*
  • Humans
  • Melanoma / pathology
  • Mice
  • Mice, Inbred C57BL
  • Mitogen-Activated Protein Kinase Kinases / antagonists & inhibitors
  • Protein Kinase Inhibitors / pharmacology
  • raf Kinases / antagonists & inhibitors

Substances

  • Protein Kinase Inhibitors
  • raf Kinases
  • Mitogen-Activated Protein Kinase Kinases