A simple method to identify kinases that regulate embryonic stem cell pluripotency by high-throughput inhibitor screening

Charles A. C. Williams, Nathanael S. Gray, Greg M. Findlay (Lead / Corresponding author)

Research output: Contribution to journalArticle

1 Citation (Scopus)
185 Downloads (Pure)

Abstract

Embryonic stem cells (ESCs) can self-renew or differentiate into all cell types, a phenomenon known as pluripotency. Distinct pluripotent states have been described, termed “naïve” and “primed” pluripotency. The mechanisms that control naïve-primed transition are poorly understood. In particular, we remain poorly informed about protein kinases that specify naïve and primed pluripotent states, despite increasing availability of high-quality tool compounds to probe kinase function. Here, we describe a scalable platform to perform targeted small molecule screens for kinase regulators of the naïve-primed pluripotent transition in mouse ESCs. This approach utilizes simple cell culture conditions and standard reagents, materials and equipment to uncover and validate kinase inhibitors with hitherto unappreciated effects on pluripotency. We discuss potential applications for this technology, including screening of other small molecule collections such as increasingly sophisticated kinase inhibitors and emerging libraries of epigenetic tool compounds.
Original languageEnglish
Article numbere55515
Number of pages6
JournalJoVE: Journal of Visualized Experiments
Issue number123
DOIs
Publication statusPublished - 12 May 2017

Keywords

  • Protein Kinase
  • Embryonic Stem Cell
  • Pluripotency
  • Naïve-Primed Transition
  • Kinase Inhibitors
  • High-Throughput Screen

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    Student Theses

    Identification of the ERK5 Kinase as a Key Regulator of Embryonic Stem Cell Pluripotency

    Author: Williams, C., 2017

    Supervisor: Findlay, G. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy

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