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Long-read Ribo-STAMP simultaneously measures transcription and translation at full length isoform resolution

GSE255844 Homo sapiens (human) RNA-seq (gene expression) 10 samples Submitted 2024/05/20 Platform GPL28352
At a glance

RNA-seq (gene expression) · human · 10 samples

What it is
RNA-seq (gene expression)Gene activity measured by sequencing. The summary does not mention single-cell work, so this is most likely bulk: one average per sample, the usual starting point for comparing groups.
Organism
Homo sapiens (human)
Samples
10 samples: 3 × APOBEC1-only HEK293T vs 3 × Ribo-STAMP HEK293T vs 2 × Ribo-STAMP MDA-MB-231 NT vs 2 × Ribo-STAMP MDA-MB-231 48h CoCl2Groups are read from the sample names, not from the paper. Check the list below.
Data you can get
A processed table (series matrix) and supplementary files from NCBI; the raw sequencing reads from ENA. Which of these hold counts depends on the study, so check the file list.
Paper
Long-read Ribo-STAMP simultaneously measures transcription and translation with isoform resolutionGenome research · 2024
Summary (written by the submitters)
Transcription and translation are intertwined processes where mRNA isoforms are crucial intermediaries. However, methodological limitations in analyzing translation at the mRNA isoform level have impaired our ability to comprehensively establish links between the full-length transcripts and the translatome. This has left gaps in our understanding of critical biological processes, regulatory mechanisms, and disease progression.
Read the full summary ↓
To address this, we develop an integrated computational and experimental framework called long-read Ribo-STAMP (LR-Ribo-STAMP). LR-Ribo-STAMP capitalizes on advancements in long-read sequencing and RNA-base editing-mediated technologies to simultaneously and scalably profile translation and transcription at both gene and mRNA isoform levels for the first time. In this report, we show agreement between gene-level translation profiles obtained with LR-Ribo-STAMP and those from previously validated short-read Ribo-STAMP data in unperturbed cells. At the mRNA isoform level, we show that LR-Ribo-STAMP successfully profiles translation in unperturbed cells and links mRNA isoforms and regulatory features, such as upstream ORFs (uORFs) and regulatory sequences, to translation measurements. We further demonstrate the method’s effectiveness in profiling disease models by profiling translation at gene and isoform levels in a triple-negative breast cancer cell line under normoxia and hypoxia. Here, we find that LR-Ribo-STAMP effectively delineates orthogonal transcriptional and translation shifts between conditions at gene and isoform levels. At the isoform level, LR-Ribo-STAMP uniquely identifies key regulatory elements and shifts in mRNA isoform transcription that correlate with changes in translational, providing an example of insight that can inform the development of novel therapeutics. Overall, LR-Ribo-STAMP is a significant advancement in translation methods and can have profound implications for basic research and clinical applications.
Published in
Long-read Ribo-STAMP simultaneously measures transcription and translation with isoform resolution
Jagannatha P, Tankka AT, Lorenz DA et al. · Genome research 2024 · PMID 38906680 · doi:10.1101/gr.279176.124
This dataset
Download · which file do I need?

Direct links to NCBI and ENA, no account and no request form. Pick by what you want to do: take the small processed table to look, the supplementary files to re-analyse, or the original raw data to start over.

1 · Just have a look

Processed table

The series matrix: one table of values per sample as the submitters processed it. The smallest download, and it opens in Excel or R. Start here to see what the data look like.

2 · Re-analyse it

Supplementary files

The count tables and other files the authors uploaded. Take only the samples you need, or the whole study as GSE255844_RAW.tar (it can run to tens of gigabytes).

3 · Start from the raw reads

Raw sequencing reads

The original FASTQ files, straight from the sequencer, held at ENA. Large, and you align them yourself, so only choose this if you need to re-process the study from scratch.

What the files usually look like: For RNA-seq the counts are usually a table among the supplementary files (look for 'counts', 'expression', 'FPKM' or 'TPM' in the file name). The series matrix of a sequencing study is sometimes nearly empty.

Also filed as BioProject PRJNA1076859 and SRA study SRP489890. Searching any of these in the dataset finder brings you back here.

Study design
3 × APOBEC1-only HEK293T vs 3 × Ribo-STAMP HEK293T vs 2 × Ribo-STAMP MDA-MB-231 NT vs 2 × Ribo-STAMP MDA-MB-231 48h CoCl2

Supports a between-group comparison across 10 samples.

How we read this: 4 replicated groups read from 10 sample titles; they account for 10 of them. These groups are guessed from the sample names, not taken from the paper, so check them against the sample list below before relying on them.

Samples in this study
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