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Distinct properties and functions of CTCF revealed by a rapidly inducible degron system [ChIP-Seq]

ABSTRACT: Distinct properties and functions of CTCF revealed by a rapidly inducible degron system [ChIP-Seq]

PROVIDER: PRJNA632004 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11780135.fastq.gz	Fastqsanger.gz
	SRR11780136.fastq.gz	Fastqsanger.gz
	SRR11780137.fastq.gz	Fastqsanger.gz
	SRR11780138.fastq.gz	Fastqsanger.gz
	SRR11780139.fastq.gz	Fastqsanger.gz

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Distinct properties and functions of CTCF revealed by a rapidly inducible degron system

Project description:Distinct properties and functions of CTCF revealed by a rapidly inducible degron system

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Distinct properties and functions of CTCF revealed by a rapidly inducible degron system [PRO-Seq]

Project description:Distinct properties and functions of CTCF revealed by a rapidly inducible degron system [PRO-Seq]

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Distinct properties and functions of CTCF revealed by a rapidly inducible degron system [ChIP-Seq]

Project description:We coupled an auxin-induced degron (AID) system with precision nuclear run-on (PRO-seq). Oriented CTCF motifs in gene bodies are associated with transcriptional stalling in a manner ostensibly independent of bound CTCF. CTCF binding sites display highly variable resistance to degradation, with persistent sites displaying architectural related features.

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Distinct properties and functions of CTCF revealed by a rapidly inducible degron system [PRO-Seq]

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Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation [HiChIP]

Project description:Background: CTCF is a well-established chromatin architectural protein that also plays various roles in transcriptional regulation. While CTCF biology has been extensively studied, how the domains of CTCF function to regulate transcription remains unknown. Additionally, the original auxin-inducible degron 1 (AID1) system has limitations to investigate the function of CTCF. Results: We employ an improved auxin-inducible degron technology, AID2, to facilitate the study of acute depletion of CTCF while overcoming the limitations of the previous AID system. As previously observed through the AID1 system and steady-state RNA analysis, the new AID2 system combined with SLAM-seq confirms that CTCF depletion leads to modest nascent and steady-state transcripts changes. A CTCF domain sgRNA library screening identifies the zinc finger (ZF) domain as the region within CTCF with the most functional relevance, including ZFs 1 and 10. Removal of ZFs 1 and 10 reveals genomic regions that independently require these ZFs for DNA binding and transcriptional regulation. Notably, loci regulated by either ZF1 or ZF10 exhibit unique CTCF binding motifs specific to each ZF. Conclusions: By extensively comparing the AID1 and AID2 systems for CTCF degradation in SEM cells, we confirm that AID2 degradation is superior for achieving miniAID tagged protein degradation without the limitations of the AID1 system. The model we create that combines AID2 depletion of CTCF with exogenous overexpression of CTCF mutants allows us to demonstrate how peripheral ZFs intricately orchestrate transcriptional regulation in a cellular context for the first time.

2022-12-09 | GSE201911 | GEO

Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation [RNA-Seq]

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Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation [SLAM-seq]

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Distinct properties and functions of CTCF revealed by a rapidly inducible degron system.

Project description:CCCTC-binding factor (CTCF) is a conserved zinc finger transcription factor implicated in a wide range of functions, including genome organization, transcription activation, and elongation. To explore the basis for CTCF functional diversity, we coupled an auxin-induced degron system with precision nuclear run-on. Unexpectedly, oriented CTCF motifs in gene bodies are associated with transcriptional stalling in a manner independent of bound CTCF. Moreover, CTCF at different binding sites (CBSs) displays highly variable resistance to degradation. Motif sequence does not significantly predict degradation behavior, but location at chromatin boundaries and chromatin loop anchors, as well as co-occupancy with cohesin, are associated with delayed degradation. Single-molecule tracking experiments link chromatin residence time to CTCF degradation kinetics, which has ramifications regarding architectural CTCF functions. Our study highlights the heterogeneity of CBSs, uncovers properties specific to architecturally important CBSs, and provides insights into the basic processes of genome organization and transcription regulation.

| S-EPMC7999233 | biostudies-literature

Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation

Project description:Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation

| PRJNA844097 | ENA

Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation [HiChIP]

Project description:Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation [HiChIP]

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