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

TitleCombinatorial knockout of RARα, RARβ, and RARγ completely abrogates transcriptional responses to retinoic acid in murine embryonic stem cells.
Publication TypeJournal Article
Year of Publication2018
AuthorsLaursen KB, Gudas LJ
JournalJ Biol Chem
Volume293
Issue30
Pagination11891-11900
Date Published2018 07 27
ISSN1083-351X
KeywordsAnimals, CRISPR-Cas Systems, Frameshift Mutation, Gene Knockout Techniques, Mice, Mouse Embryonic Stem Cells, Receptors, Retinoic Acid, Retinoic Acid Receptor alpha, Transcriptional Activation, Transcriptome, Tretinoin
Abstract

All--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.

DOI10.1074/jbc.RA118.001951
Alternate JournalJ. Biol. Chem.
PubMed ID29848550
PubMed Central IDPMC6066298
Grant ListR01 CA043796 / CA / NCI NIH HHS / United States
R01 DE010389 / DE / NIDCR NIH HHS / United States