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Development of novel integrated sequencing methods to explore translation and its regulatory mechanisms in single cells

Project description

Novel sequencing methods to study translation machinery in single cells

Recent developments in novel single-cell sequencing methods made possible the quantification and analysis of the transcriptomes in millions of individual cells. On the contrary, no current methods allow the evaluation of translation efficiencies in single cells and their correlation with the tRNA levels and modifications, including RNA-bound proteins and RNA methylation. The EU-funded scTranslatomics project aims to develop innovative technologies that will enable the investigation of translation and translation regulatory mechanisms at single-cell resolution. The novel multi-omics approaches to quantify and evaluate translation in single cells will provide information about ribosome position along the transcript, the RNA methylation status of the transcript, RNA-bound proteins, tRNA expression levels and modifications.

Objective

In recent years novel single-cell sequencing methods have allowed an in-depth analysis of the diversity of cell types and states in a wide range of organisms. Due to the continuous optimization of experimental and computational methods by many research groups, it is now possible to sequence the transcriptomes of thousands to millions of individual cells. Albeit an exciting development, transcription only covers the first step in the central dogma. The second step, the process of translation, is currently much harder to explore in single cells. Building upon existing ribosome profiling protocols, our laboratory recently majorly increased the sensitivity of these assays allowing ribosome profiling in single cells. However, currently no methods exist to determine the translation efficiencies in single cells and to correlate translation efficiencies to tRNA levels and their modifications, RNA bound proteins, and m6A methylation of mRNA, all major regulatory mechanisms of translation. The overarching goal of this proposal is to develop several novel multi-omics approaches to quantify translation in single cells by integrating information on ribosome position along the transcript, tRNA expression levels, tRNA modifications, RNA-bound proteins, and the m6A methylation status of the transcript. These technologies will open novel avenues to start exploring translation and its regulatory mechanisms with single-cell resolution. Whereas major discoveries have been made during the last decade by exploring the genome, the epigenome, and the transcriptome using single-cell sequencing approaches, translation, the major second step in the central dogma, is still very much unexplored at the single-cell level. I hope that the tools, presented in this proposal will provide the community with methods to explore, and ultimately understand, how translation contributes to the astonishing heterogeneity among single cells.

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HORIZON-ERC - HORIZON ERC Grants

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Call for proposal

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(opens in new window) ERC-2021-ADG

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Host institution

KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN - KNAW
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 2 500 000,00
Address
KLOVENIERSBURGWAL 29 HET TRIPPENHUIS
1011 JV AMSTERDAM
Netherlands

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Total cost

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€ 2 500 000,00

Beneficiaries (1)

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