Ribosomal profiling with cellular specificity

Full Project Title

Ribosomal profiling with cellular specificity to understand neuromuscular remodelling.

Project Summary

Subcellular transcriptomics remains highly expensive and lacks the resolution to assess transcripts at the level of the ribosome in specific cellular populations. Additionally, large discrepancies exist between RNA sequencing and protein expression data sets suggesting a disconnect in the way the field integrates omics-based data.  

Ribosome profiling allows for the analysis of mRNAs located at the ribosome and provides a far more accurate assessment of translational activity compared to bulk or whole cell sequencing methodologies. The aim of this project is to create a broadly accessible pipeline for UQ researchers to undertake transcriptional profiling of samples specifically enriched for ribosomal material, in their tissue of interest, using the innovative combination of adeno-associated viral vectors (AAVs), transgenic mice and established RNA sequencing methodologies, with the potential for transferability to a variety of mouse models and in vitro cell culture systems.

Potential Outcomes

An important outcome from this project will be the development of a ribosome profiling pipeline enabling UQ researchers to undertake ribosome-specific RNA sequencing in any tissue or cell line that can be manipulated with AAVs and Cre recombinase technology, with detailed and validated methodologies for performing these experiments.

Additionally, the project will provide molecular biology tools for performing this work which can be made available to other UQ researchers. Importantly, to allow flexibility and broad applicability, these tools will be developed in a modular fashion, allowing UQ researchers to “plug and play” specific components to create constructs designed to facilitate ribosome profiling in their tissue or cells of interest.