Experiment / E80WHN1H43' UTR / RNA Stability MPRA (MPRAu)

Tetracycline-responsive hammerhead riboswitch screen

High-throughput identification of synthetic riboswitches by barcode-free amplicon-sequencing in human cells

An episomal CMV-eGFP 3′-UTR library containing all 16,384 combinations of a seven-nucleotide communication module was screened in HEK-293H cells. Self-barcoding cDNA amplicon counts were measured after 3 h untreated, 25 µM, or 50 µM tetracycline stimulation, with eight biological replicates per condition.

Processed tables are specific to each experiment. Column names, units, measurements, and table structure are not standardized across the database. Check this experiment’s column definitions and quality-control notes before comparing or combining data.

Perturbation & assay details

Untreated, 25 µM tetracycline, or 50 µM tetracycline for 3 h

Transient episomal plasmid reporter with the synthetic riboswitch library in the eGFP 3′ UTR. Each construct is identified by its sequence rather than an external barcode; cDNA amplicon abundance was sequenced on an Illumina NextSeq 500, while the published log2 fold changes and FDRs were obtained from edgeR analysis of the source-data workbook.

Processed data

50 rows per page. Click a cell to inspect its full value.

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Filters apply to this table only. The CSV download contains the complete processed table; filtered rows are available through the API.

Column dictionary · 42 definitions
variant_id
Unique identifier combining the amplicon pattern name and the two variable sequence parts.
pattern_name
Amplicon sequence pattern; Orig denotes the Tet-hammerhead library.
variable_region_1
First variable sequence segment of the self-barcoding amplicon.
variable_region_2
Second variable sequence segment of the self-barcoding amplicon.
motif
The two variable sequence segments joined with an underscore.
total_count
Sum of raw cDNA amplicon abundance counts across all 24 screen samples.
min_detected_replicates_per_condition
Smallest number of nonzero-count replicates among untreated, 25 µM, and 50 µM tetracycline conditions.
count_untreated_rep1
Raw cDNA amplicon count for untreated biological replicate 1 (GSM4259904).
count_untreated_rep2
Raw cDNA amplicon count for untreated biological replicate 2 (GSM4259905).
count_untreated_rep3
Raw cDNA amplicon count for untreated biological replicate 3 (GSM4259906).
count_untreated_rep4
Raw cDNA amplicon count for untreated biological replicate 4 (GSM4259907).
count_untreated_rep5
Raw cDNA amplicon count for untreated biological replicate 5 (GSM4259908).
count_untreated_rep6
Raw cDNA amplicon count for untreated biological replicate 6 (GSM4259909).
count_untreated_rep7
Raw cDNA amplicon count for untreated biological replicate 7 (GSM4259910).
count_untreated_rep8
Raw cDNA amplicon count for untreated biological replicate 8 (GSM4259911).
count_tet_25uM_rep1
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 1 (GSM4259912).
count_tet_25uM_rep2
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 2 (GSM4259913).
count_tet_25uM_rep3
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 3 (GSM4259914).
count_tet_25uM_rep4
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 4 (GSM4259915).
count_tet_25uM_rep5
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 5 (GSM4259916).
count_tet_25uM_rep6
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 6 (GSM4259917).
count_tet_25uM_rep7
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 7 (GSM4259918).
count_tet_25uM_rep8
Raw cDNA amplicon count for 25 µM tetracycline biological replicate 8 (GSM4259919).
count_tet_50uM_rep1
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 1 (GSM4259920).
count_tet_50uM_rep2
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 2 (GSM4259921).
count_tet_50uM_rep3
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 3 (GSM4259922).
count_tet_50uM_rep4
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 4 (GSM4259923).
count_tet_50uM_rep5
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 5 (GSM4259924).
count_tet_50uM_rep6
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 6 (GSM4259925).
count_tet_50uM_rep7
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 7 (GSM4259926).
count_tet_50uM_rep8
Raw cDNA amplicon count for 50 µM tetracycline biological replicate 8 (GSM4259927).
mean_cpm_untreated
Mean per-sample counts-per-million for the untreated replicates, normalized within the Orig library.
mean_cpm_tet_25uM
Mean per-sample counts-per-million for the 25 µM tetracycline replicates, normalized within the Orig library.
mean_cpm_tet_50uM
Mean per-sample counts-per-million for the 50 µM tetracycline replicates, normalized within the Orig library.
log2fc_tet_25uM_vs_untreated
Published edgeR log2 fold change for 25 µM tetracycline versus untreated.
fdr_tet_25uM_vs_untreated
Published Benjamini–Hochberg FDR for the 25 µM versus untreated comparison.
significant_tet_25uM_vs_untreated_fdr_0_05
True when the published FDR for 25 µM versus untreated is ≤ 0.05.
log2fc_tet_50uM_vs_untreated
Published edgeR log2 fold change for 50 µM tetracycline versus untreated.
fdr_tet_50uM_vs_untreated
Published Benjamini–Hochberg FDR for the 50 µM versus untreated comparison.
significant_tet_50uM_vs_untreated_fdr_0_05
True when the published FDR for 50 µM versus untreated is ≤ 0.05.
source_stats_sheet
Source-data workbook sheet supplying the published statistics; 2d.
source_stats_available
Indicates that published log2FC and FDR values are available for the row.

Quality control

The authors assessed library abundance, replicate concordance, GC dependence, and low-abundance behavior in the accompanying pipeline and supplementary information. For this package, missing construct counts were treated as zero; a construct was retained only when it had total count > 0, was detected in at least 2 of 8 replicates in every condition, and had finite published log2FC and FDR for both tetracycline contrasts. All 16,384 input constructs passed these filters.

Curation notes

The raw GEO archive includes separate library-QC samples (GSM4259928–GSM4259932); they are preserved in raw_data but intentionally excluded from this screen table. The GEO SOFT metadata contains a treatment-label inconsistency for the final eight samples; sample titles and the paper’s stated 8+8+8 design were used to assign the 50 µM condition. The library is fully synthetic, so no reference genome or genomic interval applies.

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