Experiment / E6X1WPCCJ5' UTR / Translation Efficiency MPRA (MPTA)

eGFP random 50-mer library, N1-methylpseudouridine

Human 5′ UTR design and variant effect prediction from a massively parallel translation assay

Duplicate polysome-profiling measurements of a 280,000-member random 50-nt 5′-UTR library in a constant eGFP reporter context using N1-methylpseudouridine-substituted IVT mRNA in HEK293T cells. The processed table contains sequence-supported replicate MRLs and a derived two-replicate mean.

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

N1-methylpseudouridine (m1Ψ) substituted for uridine in IVT mRNA

HEK293T cells were transfected with in-vitro-transcribed reporter mRNA; cells were harvested 12 h later, polysome fractions were collected, fraction-associated sequences were quantified by barcode/NGS, and Mean Ribosome Load (MRL) was computed from relative fraction distributions. The reporter used a defined 25-nt leader and constant eGFP CDS/3′ context; this is an RNA polysome-profiling MPTA rather than a DNA plasmid MPRA. GEO samples GSM3130439 and GSM3130440 are the two biological replicates.

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 · 12 definitions
element_id_rep1
Raw GEO row identifier for the first biological replicate.
element_id_rep2
Raw GEO row identifier for the second biological replicate.
utr_sequence
Uppercase tested 5′-UTR insert sequence; fixed reporter CDS overlap removed where present.
utr_length
Length of the cleaned 5′-UTR insert in nucleotides.
uaug_count
Number of ATG motifs in the cleaned insert; reading frame is not inferred.
coverage_metric_rep1
GEO-provided per-element coverage metric used for within-replicate ranking; source field is total for this experiment.
coverage_metric_rep2
GEO-provided per-element coverage metric used for within-replicate ranking; source field is total for this experiment.
mrl_rep1
Published mean ribosome load (MRL) for replicate 1.
mrl_rep2
Published mean ribosome load (MRL) for replicate 2.
mrl_mean
Arithmetic mean of the two published replicate MRL values.
mrl_sd
Sample standard deviation of the two replicate MRL values.
mrl_delta_rep2_minus_rep1
Replicate 2 MRL minus replicate 1 MRL.

Quality control

Applied the authors’ sequence-processing rule: valid reporter sequence and the top 280,000 eGFP members ranked by the GEO-provided coverage field per replicate. For this generated table, additionally required A/C/G/T sequence, an expected 50-nt insert, positive finite coverage metric and finite MRL in both replicates, collapsed duplicate cleaned sequences to the highest-coverage row, and retained only sequences observed in both QC-passed replicates (269,304 rows). The raw GEO fraction fields remain in GSE114002_RAW.tar.

Curation notes

GSM3130439 contains an inconsistent first fraction column (header 00 but integer-like values), 1,391 missing rl rows, and the source total field is normalized signal. GSM3130440 has a 4-nt eGFP overlap and 1,802 missing rl rows. Invalid or missing-MRL rows were excluded; source rl was used unchanged. The simple MRL summaries are derived, and coverage_metric values are not assumed to be comparable read counts.

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