HTS-Compatible CometChip Enables Genetic Screening for Modulators of Apoptosis and DNA Double-Strand Break Repair.

Published

Journal Article

Dysfunction of apoptosis and DNA damage response pathways often drive cancer, and so a better understanding of these pathways can contribute to new cancer therapeutic strategies. Diverse discovery approaches have identified many apoptosis regulators, DNA damage response, and DNA damage repair proteins; however, many of these approaches rely on indirect detection of DNA damage. Here, we describe a novel discovery platform based on the comet assay that leverages previous technical advances in assay precision by incorporating high-throughput robotics. The high-throughput screening (HTS) CometChip is the first high-throughput-compatible assay that can directly detect physical damage in DNA. We focused on DNA double-strand breaks (DSBs) and utilized our HTS CometChip technology to perform a first-of-its-kind screen using an shRNA library targeting 2564 cancer-relevant genes. Conditions of the assay enable detection of DNA fragmentation from both exogenous (ionizing radiation) and endogenous (apoptosis) sources. Using this approach, we identified LATS2 as a novel DNA repair factor as well as a modulator of apoptosis. We conclude that the HTS CometChip is an effective assay for HTS to identify modulators of physical DNA damage and repair.

Full Text

Duke Authors

Cited Authors

  • Tay, IJ; Park, JJH; Price, AL; Engelward, BP; Floyd, SR

Published Date

  • September 2020

Published In

Volume / Issue

  • 25 / 8

Start / End Page

  • 906 - 922

PubMed ID

  • 32452708

Pubmed Central ID

  • 32452708

Electronic International Standard Serial Number (EISSN)

  • 2472-5560

Digital Object Identifier (DOI)

  • 10.1177/2472555220918367

Language

  • eng

Conference Location

  • United States