Project description:We performed ribosome profiling of polysomes (Poly-RIBOseq) to monitor translation of RNAs associated with polysomes in brain extracts from a mouse model Keywords: ribosome profiling, translation, polysome profiling, Poly-RIBOseq
Project description: Ribosome profiling (RiboSeq) has emerged as a powerful technique for studying the genome-wide regulation of translation in various cells. Several steps in the biological protocol have been improved, but the bioinformatics part of RiboSeq suffers from a lack of standardization, preventing the straightforward and complete reproduction of published results. Too many published studies provide insufficient detail about the bioinformatics pipeline used. The broad range of questions that can be asked with RiboSeq makes it difficult to use a single bioinformatics tool. Indeed, many scripts have been published for addressing diverse questions. Here, we propose a unique tool (for use with multiple operating systems, OS) to standardize the general steps that must be performed systematically in RiboSeq analysis, together with the statistical analysis and quality control of the sample. The data generated can then be exploited with more specific tools. We hope that this tool will help to standardize bioinformatics analyses pipelines in the field of translation.
Project description:blanc09_ripseq_rfl8-riboseq analysis of the rfl8 mutant-Which mitochondrial transcripts are bound by RFL8 protein? -Does the RFL8 loss impact on mitochondrial translation?
Project description:Here we applied RNAseq and RIBOseq, the deep sequencing of ribosome-associated transcripts, to investigate the relation of RNA abundance and translation at four stages of neurogenic differentiation.
Project description:We perform Ribosome Profiling (Riboseq) analysis of mouse Neuro2a neuronal cultures in Ebp1-siRNA knockdown vs. scrambled-siRNA control conditions in biological triplicate to assess the translation-specific function of Ebp1
Project description:We show that ribosome collisions, induced by Rptor deletion or aminoacid starvation, causes a ZAK alpha mediated identity switch in intestinal stem cells by activating a more fetal-like stem cell signature. This switch also causes changes in metabolic profiles of the cells, which is heavily regulated by RNA translation efficiency of different group of metabolic genes. These observations originate from performing in vitro Riboseq and RNAseq in mouse intestinal organoids, and in vivo Riboseq in Lgr5 expressing intestinal stem cells from WT and Rptor.fl/fl mice.
Project description:To identify DDX3X-dependent translation targets in the developing mouse cortex, we used E11.5 cortices from both sexes of wildtype and conditional Ddx3x knockout mice (driven by Emx1-Cre). We performed RNAseq and Riboseq on these samples in parallel.
Project description:Ribosome profiling (RiboSeq) analysis of murine 17 clone 1 (17Cl-1) cells with and without Tunicamycin treatment. Tunicamycin is known to induce the unfolded protein response, and the objective of this work was to assess the impact of Tunicamycin on cellular translation. Additionally, we sought to assess the impact of differing library preparation methods by using three separate approaches: flash freezing, 1X Cycloheximide, and 100X Cycloheximide.
Project description:We optimized a polysome profiling protocol considering the unique morphology and ribosomal distribution of astrocytes. This ‘astrocyte-optimized’ protocol was then used to study primary astrocytes treated with proinflammatory cytokines, enabeling Riboseq analysis for measurment of the transcriptome, translatome, and translation efficiency with a coverage of over 12,000 genes. We found several regulation strategies of gene expression in astrocytes executed as part of an extensive inflamatory response. Enhanced translation efficiency was one of these strategies, used specifically for genes related to oxidative phosphorylation, and ribosomal proteins.
