Liu et al. 2019 (PRJNA477831)

General Details

Title Optimization of Ribosome Profiling Using Brain Tissue from Fragile X Mice
Organism
Number of Samples 37
Release Date 2018/06/25 00:00
Sequencing Types
Protocol Details

Study Links

Repository Details

SRA SRP151279
ENA SRP151279
GEO
BioProject PRJNA477831

Publication

Title
Authors Liu B,Molinaro G,Shu H,Stackpole EE,Huber KM,Richter JD
Journal Nucleic acids research
Publication Date 2019 Mar 18
Abstract Dysregulated protein synthesis is a major underlying cause of many neurodevelopmental diseases including fragile X syndrome. In order to capture subtle but biologically significant differences in translation in these disorders, a robust technique is required. One powerful tool to study translational control is ribosome profiling, which is based on deep sequencing of mRNA fragments protected from ribonuclease (RNase) digestion by ribosomes. However, this approach has been mainly applied to rapidly dividing cells where translation is active and large amounts of starting material are readily available. The application of ribosome profiling to low-input brain tissue where translation is modest and gene expression changes between genotypes are expected to be small has not been carefully evaluated. Using hippocampal tissue from wide type and fragile X mental retardation 1 (Fmr1) knockout mice, we show that variable RNase digestion can lead to significant sample batch effects. We also establish GC content and ribosome footprint length as quality control metrics for RNase digestion. We performed RNase titration experiments for low-input samples to identify optimal conditions for this critical step that is often improperly conducted. Our data reveal that optimal RNase digestion is essential to ensure high quality and reproducibility of ribosome profiling for low-input brain tissue. © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.
PMC PMC6411937
PMID 30590705
DOI
Run Accession Study Accession Scientific Name Cell Line Library Type Treatment GWIPS-viz Trips-Viz Reads BAM BigWig (F) BigWig (R)
SRR7883717 PRJNA477831 Mus musculus Ribo-Seq
SRR7883718 PRJNA477831 Mus musculus Ribo-Seq
SRR7883719 PRJNA477831 Mus musculus Ribo-Seq
SRR7883720 PRJNA477831 Mus musculus Ribo-Seq
SRR7883721 PRJNA477831 Mus musculus Ribo-Seq
SRR7883722 PRJNA477831 Mus musculus Ribo-Seq
SRR7883723 PRJNA477831 Mus musculus Ribo-Seq
SRR7883724 PRJNA477831 Mus musculus Ribo-Seq
SRR7883725 PRJNA477831 Mus musculus Ribo-Seq
SRR7883726 PRJNA477831 Mus musculus Ribo-Seq
SRR7883727 PRJNA477831 Mus musculus Ribo-Seq
SRR7883728 PRJNA477831 Mus musculus Ribo-Seq
SRR7883729 PRJNA477831 Mus musculus Ribo-Seq
SRR7883730 PRJNA477831 Mus musculus Ribo-Seq
SRR7883731 PRJNA477831 Mus musculus Ribo-Seq
SRR7883746 PRJNA477831 Mus musculus Ribo-Seq
SRR7883747 PRJNA477831 Mus musculus Ribo-Seq
SRR7883748 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883749 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883750 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883751 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883752 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883753 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883754 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883755 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883756 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883757 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883758 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883759 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883760 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883761 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883762 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883763 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883764 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883765 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883766 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
SRR7883767 PRJNA477831 Homo sapiens iPSC Ribo-Seq Cycloheximide
Run Accession Study Accession Scientific Name Cell Line Library Type Treatment GWIPS-viz Trips-Viz Reads BAM BigWig (F) BigWig (R)

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