Experiment / E3IFSCAGD5' UTR / Translation Efficiency MPRA (MPTA)

PC3 small PLUMAGE 8-bp barcode proof-of-principle

Multiplexed functional genomic analysis of 5′ untranslated region mutations across the spectrum of prostate cancer

A small episomal PLUMAGE library tested full-length wild-type and mutant 5′-UTRs from ADAM32, COMT, and ZCCHC7 in PC3 cells. Each construct was linked to five known 8-bp barcodes; the packaged table combines PacBio construct-linkage counts with the published transcript and translation-efficiency summaries and pilot replicate values.

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

Basal / Untreated (24 h post-transfection)

Episomal pGL3-promoter luciferase reporter constructs carried full-length human 5′-UTRs and a fixed, known 8-bp barcode cloned at the 3′ end of the luciferase coding sequence. The pooled library was transfected into PC3 cells; DNA, total mRNA, and polysome-bound mRNA were sequenced, and translation efficiency was measured as normalized polysome signal relative to normalized total-RNA signal.

Processed data

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

Visible columns (28 of 28)
Row
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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 · 28 definitions
variant_id
Stable variant identifier using the gene, hg19 chromosome, position, reference allele, and alternative allele.
gene
Gene symbol for the assayed 5′-UTR.
variant_position_hg19
hg19 genomic position of the tested 5′-UTR mutation.
reference_allele
Reference nucleotide at the tested position.
alternative_allele
Mutant nucleotide at the tested position.
cell_line
Cell line used for the pilot assay.
barcode_design
Description of the fixed barcode design and its relationship to the WT and mutant constructs.
wild_type_barcodes
Semicolon-separated known 8-bp barcodes assigned to the WT construct.
mutant_barcodes
Semicolon-separated known 8-bp barcodes assigned to the mutant construct.
wild_type_barcode_count
Number of WT barcodes retained in the table.
mutant_barcode_count
Number of mutant barcodes retained in the table.
wild_type_total_long_read_count
Sum of author-reported PacBio long-read counts across WT barcodes.
mutant_total_long_read_count
Sum of author-reported PacBio long-read counts across mutant barcodes.
wild_type_mean_long_read_count
Mean author-reported PacBio long-read count per WT barcode.
mutant_mean_long_read_count
Mean author-reported PacBio long-read count per mutant barcode.
wild_type_translation_efficiency_replicates
Semicolon-separated pilot normalized polysome/total-RNA values for individual WT barcodes from Source Data Figure 1g.
mutant_translation_efficiency_replicates
Semicolon-separated pilot normalized polysome/total-RNA values for individual mutant barcodes from Source Data Figure 1g.
transcript_log2_fold_change
Published log2 fold change for mutant versus WT transcript activity from the PLUMAGE summary.
transcript_p_value
Published two-sided Mann–Whitney U-test p-value for the transcript comparison.
transcript_fdr
Published FDR-adjusted p-value for the transcript comparison.
transcript_significant_fdr_lt_0_1
Whether the published transcript FDR is below 0.1.
translation_efficiency_log2_fold_change
Published log2 fold change for mutant versus WT translation efficiency; blank means no corresponding row was supplied in Supplementary Data 6e.
translation_efficiency_p_value
Published p-value for the translation-efficiency comparison; blank means unavailable in the supplied summary.
translation_efficiency_fdr
Published FDR-adjusted p-value for the translation-efficiency comparison; blank means unavailable in the supplied summary.
translation_efficiency_significant_fdr_lt_0_1
Whether the published translation-efficiency FDR is below 0.1; false when no published summary row is available.
functional_in_any_layer
Whether the published FDR is below 0.1 in either the transcript or translation-efficiency layer.
qc_pass
Package-level QC flag; only true rows are present in the processed table.
source_data
Supplementary workbooks and source-data sheet used to construct the row.

Quality control

The paper reports that all 30 pilot 8-bp barcodes were detected and linked to their expected full-length WT or mutant 5′-UTR by long-read sequencing. For this package, a row passed QC only when both alleles had exactly five unique barcodes and every packaged long-read count was positive; all three variant pairs passed, so no candidate row was removed. Published FDR values are retained as effect annotations rather than used to discard nonsignificant variants.

Curation notes

PC3 corresponds to Cellosaurus CVCL:0035. Supplementary Data 6e does not provide a ZCCHC7_MUT_8mer effect-summary row, so its translation-efficiency summary fields are blank rather than treated as zero or nonsignificant; the available ZCCHC7 pilot replicate values from Source Data Figure 1g are retained. The five-barcode counts are construct-linkage/long-read counts and should not be interpreted as RNA/DNA activity measurements.

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