Impact of RNA modifications on neuronal fate

Project Information

Rn7SKmod

Grant agreement ID: 896324

DOI

10.3030/896324

Project closed

EC signature date 24 March 2020

Start date 1 July 2021

End date 30 June 2023

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 162 806,40

EU contribution

€ 162 806,40

Coordinated by

DEUTSCHES KREBSFORSCHUNGSZENTRUM HEIDELBERG
Germany

Periodic Reporting for period 1 - Rn7SKmod (Impact of RNA modifications on neuronal fate)

Reporting period: 2021-07-01 to 2023-06-30

Summary of the context and overall objectives of the project

The project aimed to investigate the functional roles of RNA modifications within the non-coding RNA Rn7sk, with a specific focus on their involvement in regulating neural fate decisions. The objectives of the project were threefold:

1. Identification and modulation of Rn7sk RNA modifications: The first objective (WP 1) was to comprehensively identify all RNA modifications specific to Rn7sk and subsequently examine how these modifications influence interactions between RNA and proteins. By delving into this aspect, the project sought to uncover the intricate mechanisms through which RNA modifications shape vital cellular processes.

2. Neuronal differentiation regulation: The second objective (WP 2) revolved around unraveling the mechanisms by which Rn7sk contributes to the regulation of neuronal differentiation in mouse embryonic stem cells (mESCs). This aspect of the study aimed to shed light on the pivotal role of Rn7sk in steering the development of neural cells and pathways.

3. Transcriptional impact of Rn7sk-deletion: The third objective (WP 3) involved analyzing the transcriptional effects arising from the deletion of Rn7sk within the forebrain. By examining the changes in gene expression patterns resulting from this deletion, the project aimed to provide insights into the broader consequences of Rn7sk on brain development and function.

The significance of this project extended to society at large due to its potential to uncover crucial insights into brain development and function. By understanding how RNA modifications influence neural fate decisions, the research had the potential to offer groundbreaking perspectives on neurodevelopmental disorders. These findings could lead to the identification of novel therapeutic targets, thereby paving the way for clinical interventions that address the complexities of such disorders.

In conclusion, this MSCA European individual postdoc fellowship centered around investigating the impact of RNA modifications within Rn7sk on neural fate decisions. Through comprehensive exploration and analysis, the project aimed to contribute to the understanding of brain development and its potential applications in tackling neurodevelopmental disorders. The insights gained from the project have the potential to open new avenues for therapeutic interventions and offer hope for individuals facing such challenges.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

Initially, I tried hybridization-based methods for six months to identify RNA modifications in Rn7sk using mass spectrometry. However, yields were consistently too low. Collaborating with Sylvia Erhard's group, we switched to using nanoswitches to obtain sufficient amount of pure Rn7sk RNA for mass spectrometry. This was successful and samples are being analyzed by Stefanie Kellner's lab. Concurrently, I'm setting up a nanopore sequencing protocol for direct RNA-modification analysis as an alternative method for determining Rn7sk RNA modifications.

As detailed in the Periodic Technical Report, Part B, section 5, my project shifted from the original objectives due to negative data, group relocation, and COVID-19 restrictions. This prompted alternative approaches to study RNA modifications' impact on gene expression. The results led to a co-first authorship in a Nature manuscript, currently in revision.

During the reporting period, research findings were communicated locally through oral presentations at the Frye Laboratory retreat in 2020 and the Single Cell Center Heidelberg symposium in 2021. Globally, results were shared via posters at the Gordon Research Conference on Epithelia Differentiation and Keratinization in 2019 and 2023. A manuscript is being revised for publication, and once accepted, the results will be shared on social media platforms, including X (formerly Twitter) or similar.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

This project's outcomes have unveiled groundbreaking insights into gene expression in a non-model organism that resists neurodevelopmental and age-related diseases, holding the potential to inspire new strategies for disease prevention and intervention, thereby contributing to improved societal well-being.

Preliminary data Fig.1