An isogenic cell line panel for sequence-based screening of targeted anticancer drugs

Ashley L. Cook, Nicolas Wyhs, Surojit Sur, Blair Ptak, Maria Popoli, Laura Dobbyn, Tasos Papadopoulos, Chetan Bettegowda, Nickolas Papadopoulos, Bert Vogelstein, Shibin Zhou, Kenneth W. Kinzler

Research output: Contribution to journalArticlepeer-review

Abstract

We describe the creation of an isogenic cell line panel representing common cancer pathways, with features optimized for high-throughput screening. More than 1,800 cell lines from three normal human cell lines were generated using CRISPR technologies. Surprisingly, most of these lines did not result in complete gene inactivation despite integration of sgRNA at the desired genomic site. A subset of the lines harbored biallelic disruptions of the targeted tumor suppressor gene, yielding a final panel of 100 well-characterized lines covering 19 frequently lost cancer pathways. This panel included genetic markers optimized for sequence-based ratiometric assays for drug-based screening assays. To illustrate the potential utility of this panel, we developed a high-throughput screen that identified Wee1 inhibitor MK-1775 as a selective growth inhibitor of cells with inactivation of TP53. These cell lines and screening approach should prove useful for researchers studying a variety of cellular and biochemical phenomena.

Original languageEnglish (US)
Article number104437
JournaliScience
Volume25
Issue number6
DOIs
StatePublished - Jun 17 2022

Keywords

  • Biochemical analysis
  • Biochemistry
  • Cancer

ASJC Scopus subject areas

  • General

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