Combinatorial knockout of RARα, RARβ, and RARγ completely abrogates transcriptional responses to retinoic acid in murine embryonic stem cells. Academic Article uri icon

Overview

abstract

  • All-trans-retinoic acid (RA), a potent inducer of cellular differentiation, functions as a ligand for retinoic acid receptors (RARα, β, and γ). RARs are activated by ligand binding, which induces transcription of direct genomic targets. However, whether embryonic stem cells respond to RA through routes that do not involve RARs is unknown. Here, we used CRISPR technology to introduce biallelic frameshift mutations in RARα, RARβ, and RARγ, thereby abrogating all RAR functions in murine embryonic stem cells. We then evaluated RA-responsiveness of the RAR-null cells using RNA-Seq transcriptome analysis. We found that the RAR-null cells display no changes in transcripts in response to RA, demonstrating that the RARs are essential for the regulation of all transcripts in murine embryonic stem cells in response to RA. Our key finding, that in embryonic stem cells the transcriptional effects of RA all depend on RARs, addresses a long-standing topic of discussion in the field of retinoic acid signaling.

publication date

  • May 30, 2018

Research

keywords

  • Gene Knockout Techniques
  • Mouse Embryonic Stem Cells
  • Receptors, Retinoic Acid
  • Retinoic Acid Receptor alpha
  • Transcriptome

Identity

PubMed Central ID

  • PMC6066298

Scopus Document Identifier

  • 85050741844

Digital Object Identifier (DOI)

  • 10.1038/nbt.1754

PubMed ID

  • 29848550

Additional Document Info

volume

  • 293

issue

  • 30