A protocol for rapid degradation of endogenous transcription factors in mammalian cells and identification of direct regulatory targets

Hillary M. Layden, Nicholas A. Eleuteri, Scott W. Hiebert, Kristy R. Stengel

Research output: Contribution to journalArticlepeer-review

Abstract

Transcriptional changes happen within minutes; however, RNAi or genetic deletion requires days to weeks before transcription networks can be analyzed. This limitation has made it challenging to distinguish direct from indirect targets of sequence-specific transcription factors. This inability to define direct transcriptional targets hinders detailed studies of transcriptional mechanisms. This protocol combines rapid degradation of endogenous transcription factors with nascent transcript analysis to define the earliest, and likely direct, regulatory targets of transcription factors. For complete details on the use and execution of this protocol, please refer to Stengel et al., 2021).

Original languageEnglish (US)
Article number100530
JournalSTAR Protocols
Volume2
Issue number2
DOIs
StatePublished - Jun 18 2021
Externally publishedYes

Keywords

  • Cell Biology
  • CRISPR
  • Flow Cytometry/Mass Cytometry
  • Genomics
  • Molecular Biology
  • Protein Biochemistry

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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