A novel microscopy-based high-throughput screening method to identify proteins that regulate global histone modification levels

Roy Baas, Daphne Lelieveld, Hetty Van Teeffelen, Philip Lijnzaad, Bas Castelijns, F. M. Van Schaik, Michiel Vermeulen, David A. Egan, H. Th Marc Timmers, Petra De Graaf

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Posttranslational modifications of histones play an important role in the regulation of gene expression and chromatin structure in eukaryotes. The balance between chromatin factors depositing (writers) and removing (erasers) histone marks regulates the steady-state levels of chromatin modifications. Here we describe a novel microscopy-based screening method to identify proteins that regulate histone modification levels in a high-throughput fashion. We named our method CROSS, for Chromatin Regulation Ontology SiRNA Screening. CROSS is based on an siRNA library targeting the expression of 529 proteins involved in chromatin regulation. As a proof of principle, we used CROSS to identify chromatin factors involved in histone H3 methylation on either lysine-4 or lysine-27. Furthermore, we show that CROSS can be used to identify chromatin factors that affect growth in cancer cell lines. Taken together, CROSS is a powerful method to identify the writers and erasers of novel and known chromatin marks and facilitates the identification of drugs targeting epigenetic modifications.

Original languageEnglish
Pages (from-to)287-296
Number of pages10
JournalJournal of Biomolecular Screening
Volume19
Issue number2
DOIs
Publication statusPublished - Feb 2014
Externally publishedYes

Keywords

  • cancer and cancer drugs
  • cell-based assays
  • epigenetics
  • microscopy
  • RNA interference
  • RNAi
  • shRNA

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