New dual inducible cellular model to investigate temporal control of oncogenic cooperating genes

Sci Rep. 2024 Sep 5;14(1):20773. doi: 10.1038/s41598-024-71227-3.

Abstract

The study of cooperating genes in cancer can lead to mechanistic understanding and identifying potential therapeutic targets. To facilitate these types of studies, we developed a new dual-inducible system utilizing the tetracycline- and cumate-inducible systems driving HES3 and the PAX3::FOXO1 fusion-oncogene, respectively, as cooperating genes from fusion-positive rhabdomyosarcoma. With this model, we can independently induce expression of either HES3 or PAX3::FOXO1, as well as simultaneously induce expression of both genes. This new model will allow us to further investigate the cooperation between HES3 and PAX3::FOXO1 including the temporal requirements for genetic cooperation. Functionally, we show that dual-induction of PAX3::FOXO1 and HES3 modifies sphere formation in a HEK293T-based system. More broadly, this lentiviral dual-inducible system can be adapted for any cooperating genes (overexpression or knockdown), allowing for independent, simultaneous, or temporally controlled gene expression.

Keywords: Cooperating genes; Cumate-inducible expression; Fusion-oncogene; Inducible expression; Temporal control; Tetracycline-inducible expression.

MeSH terms

  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Forkhead Box Protein O1 / genetics
  • Forkhead Box Protein O1 / metabolism
  • Gene Expression Regulation, Neoplastic*
  • HEK293 Cells
  • Humans
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism
  • PAX3 Transcription Factor / genetics
  • PAX3 Transcription Factor / metabolism
  • Rhabdomyosarcoma / genetics

Substances

  • Oncogene Proteins, Fusion
  • Basic Helix-Loop-Helix Transcription Factors
  • Forkhead Box Protein O1
  • PAX3 Transcription Factor
  • PAX3 protein, human
  • FOXO1 protein, human