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

• DOI: 10.1016/j.isci.2022.104437
• Publication Date: 2022-05-23T17:26:09Z
• Authors: Ashley L. Cook; Nicolas Wyhs; Surojit Sur; Blair Ptak; Maria Popoli; Laura Dobbyn; Tasos Papadopoulos; Chetan Bettegowda; Nickolas Papadopo...; Bert Vogelstein; Shibin Zhou; Kenneth W. Kinzler
• Keywords: High-Throughput Screening; Cancer cell lines

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