A Three-Way Combinatorial CRISPR Screen for Analyzing Interactions among Druggable Targets

Cell Rep. 2020 Aug 11;32(6):108020. doi: 10.1016/j.celrep.2020.108020.

Abstract

We present a CRISPR-based multi-gene knockout screening system and toolkits for extensible assembly of barcoded high-order combinatorial guide RNA libraries en masse. We apply this system for systematically identifying not only pairwise but also three-way synergistic therapeutic target combinations and successfully validate double- and triple-combination regimens for suppression of cancer cell growth and protection against Parkinson's disease-associated toxicity. This system overcomes the practical challenges of experimenting on a large number of high-order genetic and drug combinations and can be applied to uncover the rare synergistic interactions between druggable targets.

Keywords: CRISPR; CombiGEM; Parkinson's disease; cancer; combination therapy; combinatorial genetics; drug interaction; genetic interaction; high-throughput screening; neurodegeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • CRISPR-Cas Systems*
  • Drosophila melanogaster
  • Drug Combinations*
  • Drug Delivery Systems / methods*
  • Gene Knockout Techniques
  • HEK293 Cells
  • High-Throughput Screening Assays / methods*
  • Humans
  • Mice
  • Neoplasms / drug therapy
  • Parkinson Disease / drug therapy
  • RNA, Guide, CRISPR-Cas Systems

Substances

  • Antineoplastic Agents
  • Drug Combinations
  • RNA, Guide, CRISPR-Cas Systems