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RNAfate

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Identifying RNA fate checkpoints by resolving the high resolution spatiotemporal...

Identifying RNA fate checkpoints by resolving the high resolution spatiotemporal binding dynamics of CBC containing complexes High-throughput transcriptomic analyses in human cell lines have found that >80% of the genome is transcriptionally active. A major part of this massive genomic output is derived from RNA polymerase II (RNAPII) activity; such as,... ver más

31/03/2021

200K€

Presupuesto del proyecto: 200K€

Líder del proyecto

AARHUS UNIVERSITET No se ha especificado una descripción o un objeto social para esta compañía.

TRL 4-5

Fecha límite participación Sin fecha límite de participación.

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Financiación concedida El organismo H2020 notifico la concesión del proyecto el día 2021-03-31

MSCA-IF-2017: Individual Fellowships Objective:The goal of the Individual Fellowships is to enhance the creative and innovative potential...

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2 Participantes

200.19K€ | Lider

BIOSYSTEMS

489.60K€ | Participante

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Duración del proyecto: 35 meses Fecha Inicio: 2018-04-17
Fecha Fin: 2021-03-31

Líder del proyecto

AARHUS UNIVERSITET No se ha especificado una descripción o un objeto social para esta compañía.

TRL 4-5

Presupuesto del proyecto 200K€

Fecha límite de participación Sin fecha límite de participación.

Descripción del proyecto High-throughput transcriptomic analyses in human cell lines have found that >80% of the genome is transcriptionally active. A major part of this massive genomic output is derived from RNA polymerase II (RNAPII) activity; such as, mRNA, sn(o)RNA and long non-coding RNA. However, although these transcripts all contain 5’-m7G caps, which are common hallmarks of RNAPII-derived transcripts, their fates differ substantially as some are rapidly degraded while others remain stable and exercise diverse functions in the cell. What is the underlying mechanism? Transcript fate decisions are ultimately dictated by the proteins with which the nascent RNA associate. Central to this process is the cap-binding complex (CBC). Through its early association with the 5’-m7G cap, the CBC directs a plethora of nuclear RNA metabolic events by serving as a landing pad to recruit productive and/or destructive factors. Therefore, composition of the early RNA-protein particle plays an essential role in dictating RNA fate, and the CBC and its cofactors pose an interesting dichotomous system to study as a model for sorting mechanisms dictating RNA fate. In my project, I will delineate the spatiotemporal recruitment kinetics of selected RNA metabolic factors to identify when RNA fate decisions are made during transcription and how RNA/DNA elements contribute. To resolve the sequential loading of the CBC and its cofactors onto elongating transcripts, I will develop time course UV cross-linking and immunoprecipitation (CLIP) experiments, combining metabolic labelling of RNA, using the photoactivatable ribonucleoside analogue 4-sU, with a new and unprecedentedly high powered UV cross-linking technology employed at multiple short time increments. This will for the first time enable the study of in vivo RNA binding kinetics of RNA-binding proteins with a temporal resolution necessary to characterise co-transcriptional RNA fate decisions.

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