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Locus folding mechanisms determine modes of antigen receptor gene assembly (RNA-seq)

ABSTRACT: Locus folding mechanisms determine modes of antigen receptor gene assembly (RNA-seq)

PROVIDER: PRJNA1049876 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR27139436_1.fastq.gz	Fastqsanger.gz
	SRR27139436_2.fastq.gz	Fastqsanger.gz
	SRR27139437_1.fastq.gz	Fastqsanger.gz
	SRR27139437_2.fastq.gz	Fastqsanger.gz
	SRR27139438_1.fastq.gz	Fastqsanger.gz

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Project description:The dynamic folding of genomes regulates numerous biological processes, including antigen receptor (AgR) gene assembly. We show that, unlike other AgR loci, homotypic chromatin interactions and bi-directional chromosome looping both contribute to structuring Tcrb for efficient long-range V(D)J recombination. Inactivation of the CTCF binding element (CBE) or promoter at the most 5’Vβ segment (Trbv1) impaired loop extrusion originating locally and extending to DβJβ CBEs at the opposite end of Tcrb. Promoter or CBE mutation nearly eliminated Trbv1 contacts and decreased RAG endonuclease-mediated Trbv1 recombination. Importantly, Trbv1 rearrangement can proceed independent of substrate orientation, ruling out scanning by DβJβ-bound RAG as the sole mechanism of Vβ recombination, distinguishing it from Igh. Our data indicate that CBE-dependent generation of loops cooperates with promoter-mediated activation of chromatin to juxtapose Vβ and DβJβ segments for recombination through diffusion-based synapsis. Thus, the mechanisms that fold a genomic region can influence molecular processes occurring in that space, which may include recombination, repair, and transcriptional programming.

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