Genome editing demonstrates that the -5 kb Nanog enhancer regulates Nanog expression by modulating RNAPII initiation and/or recruitment. J Biol Chem 2021;296:100189
Date
12/19/2020Pubmed ID
33334884Pubmed Central ID
PMC7948488DOI
10.1074/jbc.RA120.015152Scopus ID
2-s2.0-85101802426 (requires institutional sign-in at Scopus site) 8 CitationsAbstract
Transcriptional enhancers have been defined by their ability to operate independent of distance and orientation in plasmid-based reporter assays of gene expression. At present, histone marks are used to identify and define enhancers but do not consider the endogenous role of an enhancer in the context of native chromatin. We employed a combination of genomic editing, single cell analyses, and sequencing approaches to investigate a Nanog-associated cis-regulatory element, which has been reported by others to be either an alternative promoter or a super-enhancer. We first demonstrate both distance and orientation independence in native chromatin, eliminating the issues raised with plasmid-based approaches. We next demonstrate that the dominant super-enhancer modulates Nanog globally and operates by recruiting and/or initiating RNA Polymerase II. Our studies have important implications to how transcriptional enhancers are defined and how they regulate gene expression.
Author List
Agrawal P, Blinka S, Pulakanti K, Reimer MH Jr, Stelloh C, Meyer AE, Rao SAuthor
Sridhar Rao MD, PhD Associate Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsCRISPR-Cas Systems
Cell Line
Enhancer Elements, Genetic
Gene Editing
Gene Expression Regulation
Mice
Mouse Embryonic Stem Cells
Nanog Homeobox Protein
RNA Polymerase II
Transcriptional Activation
