Impact of RNA modifications on neuronal fate

Descripción del proyecto

Modificaciones del ARN en el desarrollo neuronal

Los ARN no codificantes están emergiendo como reguladores fundamentales de una gran cantidad de vías moleculares sofisticadas. El proyecto Rn7SKmod, financiado con fondos europeos, se centrará en el Rn7sk, un ARN no codificante que modula la síntesis transcripcional de la ARN polimerasa II. Su objetivo es caracterizar las modificaciones del ARN en esta molécula y determinar cómo afectan al reclutamiento de proteínas acompañantes. Mediante la espectrometría de masas de ARN y la secuenciación de todo el genoma, los científicos del proyecto investigarán el efecto de estas modificaciones en el destino de las células neuronales y el desarrollo del encéfalo. Los resultados del proyecto ofrecerán datos fundamentales sobre la regulación del desarrollo neuronal y la participación de las modificaciones del ARN.

Objetivo

The Epitranscriptome is the assembly of over 170 chemical RNA modifications that play fundamental roles in the majority of cellular processes. RNA modifying enzymes and proper control of neuronal cell fate are essential for brain development and dysfunction in any of these processes results in severe neurodevelopmental disorders. To date, the role of non-coding RNAs (ncRNAs) as key regulators of sophisticated molecular pathways is becoming increasingly clear. A key regulatory ncRNA that modulates the transcriptional output of RNA polymerase II is Rn7sk. Other abundant ncRNAs, like transfer RNAs and ribosomal RNAs, are highly modified; yet the modifications occurring in Rn7sk are largely unknown. In this proposal, I aim to investigate the Rn7sk RNA modification status and the impact of these modifications on neuronal cell fate and brain development. To accomplish this, I will; i) Characterize Rn7sk RNA modifications using a highly sensitive RNA mass spectrometry approach and evaluate the impact of selected modifications in recruiting protein binding partners using stable isotope labelling by amino acids in cell culture (SILAC) based mass spectrometry; ii) Determine the biological function of selected Rn7sk modifications during neuronal differentiation using genetically manipulated embryonic cell lines; iii) Investigate the role of Rn7sk during brain development using genome-wide sequencing technologies and a forebrain-specific Rn7sk knock-out mouse model. Results stemming from this project will reveal several novel aspects about how neuronal fate is controlled by tiny RNA modifications. My ultimate ambition is to establish my own laboratory and become an independent research leader in the field of Epitranscriptomics. This proposal, with the support from the experienced Prof Frye and the excellent host institute, DKFZ, is key for me to reach my goal.