Single-cell and isoform-specific translational profiling of the mouse brain
Nature • • health
Key Points:
- Researchers developed and optimized Ribo-STAMP constructs for studying translation in mouse neurons, including cloning into AAV vectors and production of viral particles for in vivo and in vitro experiments.
- Primary cortical neurons from embryonic mice were cultured, infected with Ribo-STAMP AAVs, and subjected to various treatments to induce translation, with subsequent analyses including proximity ligation assay (PLA), immunocytochemistry, and viability assays.
- Single-cell RNA sequencing and long-read sequencing approaches were combined with a novel Python tool, MARINE, to identify RNA editing events at single-cell and isoform resolution, enabling detailed analysis of translational regulation across cell types and transcript isoforms.
- Extensive bioinformatics pipelines were established for processing sequencing data, including normalization of editing
