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GSE94460

GSE GEO
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Ribosome profiling of HEK293 cells.

Organism: Homo sapiens
Platform: GPL16791
Samples: 3
Experiment Types:
Other
Submitted: Feb 03 2017
Last Updated: May 15 2019
Status: Public on Jan 22 2018
Contact: Yiwen,,Chen (UT MD Anderson Cancer Center)

Relations

BioProject: https://www.ncbi.nlm.nih.gov/bioproject/PRJNA369756 SRA: https://www.ncbi.nlm.nih.gov/sra?term=SRP098797

Summary

The development of the ribosome profiling (ribo-seq) technique has enabled the measurement of translation at a genome-wide level. There are several variants of the ribosome profiling technique that use different translation inhibitors. The regular ribo-seq utilizes Cycloheximide (CHX), a translation elongation inhibitor to freeze all translating ribosomes. In contrast to CHX, the translation inhibitor lactimidomycin (LTM) and harringtonine (Harr) have a much stronger effect on initiating ribosomes. The use of these inhibitors allows for the global mapping of translating initiating sites (TISs) when they are coupled with ribosome profiling (TI-seq). We have developed a computational tool to detect and/or quantitatively compare translation initiation from TI-seq data, and predict novel ORFs from regular CHX-based ribo-seq data. Two replicates of CHX-based ribo-seq experiments and one Harr based ribo-seq were performed in HEK293 cells for confirming the ORFs predicted using public available data.

Overall Design

Ribosome protected fragments were extracted from HEK293 cells treated with cycloheximide or harringtonine plus cycloheximide.

Analysis (4 steps)

View Data Processing
Processing steps for GSE94460
  1. Library strategy: Ribosome profiling
  2. Fastq reads were trimmed and mapped to human rRNA sequences using bowtie allowing 2 mismatches.
  3. Unmapped reads were mapped to human genome GRCh38 with Ensembl gene annotation release 83 using STAR with parameters ‘--outSAMattributes All --outFilterMismatchNmax 2 --alignEndsType EndToEnd --outFilterIntronMotifs RemoveNoncanonicalUnannotated --alignIntronMax 20000 --outMultimapperOrder Random --outSAMmultNmax 1’
  4. Reads with mapQ=0 or NH>5 were filtered out and only primary loci are counted.

Supplementary Files (1)

GSE94460_RAW.tar Download
GEO Samples (3)

Dataset Citations (1)

SRA Experiments (3) and Runs (3)

Total: 2076 MB
SRX2536563 SRP098797 OTHER SINGLE
GSM2476121: HEK293 RPF Rep1; Homo sapiens; OTHER
Sample: SRS1956503
BioProject: PRJNA369756
BioSample: SAMN06293621
Platform: ILLUMINA
Instrument: Illumina HiSeq 2500
Organism: Homo sapiens
Sample attributes
source_name: Human embryonic kidney cell line
cell line: HEK293
Original files (1)
Human embryonic kidney cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR5227448 33781731 1722868281 748.77 1_S1_L001_R1_001.fastq.gz, SRR5227448, SRR5227448.sralite SRA
SRX2536564 SRP098797 OTHER SINGLE
GSM2476122: HEK293 RPF Rep2; Homo sapiens; OTHER
Sample: SRS1956504
BioProject: PRJNA369756
BioSample: SAMN06293633
Platform: ILLUMINA
Instrument: Illumina HiSeq 2500
Organism: Homo sapiens
Sample attributes
source_name: Human embryonic kidney cell line
cell line: HEK293
Original files (1)
Human embryonic kidney cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR5227449 28042962 1430191062 625.04 SRR5227449, SRR5227449.sralite SRA
SRX3427420 SRP098797 OTHER SINGLE
GSM2867900: HEK293 Harr; Homo sapiens; OTHER
Sample: SRS2718885
BioProject: PRJNA369756
BioSample: SAMN08106045
Platform: ILLUMINA
Instrument: Illumina HiSeq 2500
Organism: Homo sapiens
Sample attributes
source_name: HEK293 cells
cell line: HEK293
Original files (1)
HEK293 cells
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR6327777 31545576 1608824376 702.34 SRR6327777, SRR6327777.sralite SRA

Linked Publications (1)

Data Files (3)

Accession File Name Stored Type Output Type Mapping Assembly Size Download
1_S1_L001_R1_001.fastq.gz OTHER 748.8 MB link
SRR5227449 OTHER 625.0 MB link
SRR6327777 OTHER 702.3 MB link