TINC- A Method to Dissect Regulatory Complexes at Single-Locus Resolution- Reveals an Extensive Protein Complex at the Nanog Promoter

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2020 Dec 9

PMID 33296673

Citations 8

Authors

Anja S Knaupp

Monika Mohenska

Michael R Larcombe

Ethan Ford

Sue Mei Lim

Kayla Wong

Joseph Chen

Jaber Firas

Cheng Huang

Xiaodong Liu

Trung Nguyen

Yu B Y Sun

Melissa L Holmes

Pratibha Tripathi

Jahnvi Pflueger

Fernando J Rossello

Jan Schroder

Kathryn C Davidson

Christian M Nefzger

Partha P Das

Jody J Haigh

Ryan Lister

Ralf B Schittenhelm

Jose M Polo

Affiliations

Soon will be listed here.

Abstract

Cellular identity is ultimately dictated by the interaction of transcription factors with regulatory elements (REs) to control gene expression. Advances in epigenome profiling techniques have significantly increased our understanding of cell-specific utilization of REs. However, it remains difficult to dissect the majority of factors that interact with these REs due to the lack of appropriate techniques. Therefore, we developed TINC: TALE-mediated isolation of nuclear chromatin. Using this new method, we interrogated the protein complex formed at the Nanog promoter in embryonic stem cells (ESCs) and identified many known and previously unknown interactors, including RCOR2. Further interrogation of the role of RCOR2 in ESCs revealed its involvement in the repression of lineage genes and the fine-tuning of pluripotency genes. Consequently, using the Nanog promoter as a paradigm, we demonstrated the power of TINC to provide insight into the molecular makeup of specific transcriptional complexes at individual REs as well as into cellular identity control in general.

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