Experiment / E2K2UK0FU3' UTR / RNA Stability MPRA (MPRAu)

HeLa PTRE-seq multi-fraction synthetic 3′UTR library

PTRE-seq reveals mechanism and interactions of RNA binding proteins and miRNAs

An episomal CMV/TO-EGFP reporter library containing 642 synthetic 3′UTR identities, each represented by barcoded oligonucleotides, was transfected into HeLa cells. Total RNA, polysome-associated RNA, and 40S-associated RNA were quantified against plasmid input to measure RNA expression, translation efficiency, translation initiation, and 40S association.

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

Standard episomal plasmid MPRA configuration: synthetic 3′UTR elements were cloned downstream of a CMV/TO-driven EGFP reporter and each insert was linked to a co-transcribed 9-bp barcode. HeLa cells were harvested 40 h after electroporation; total RNA and cycloheximide-stabilized 40S/polysome fractions were barcode-sequenced. The source analysis normalized barcode counts to the mean of two plasmid-input replicates and to the median 4×Blank control.

Processed data

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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 · 51 definitions
construct_id
Canonical synthetic 3′UTR reporter identity from the source library.
construct_class
Curated class of the reporter identity: blank control, synthetic element combination, let-7 variant, natural let-7 target, or let-7 copy-number variant.
element_position_1
Regulatory element at synthetic 3′UTR position 1; blank for special variant/natural-target identities whose positional encoding is not explicit in the identity label.
element_position_2
Regulatory element at synthetic 3′UTR position 2; blank for special variant/natural-target identities whose positional encoding is not explicit in the identity label.
element_position_3
Regulatory element at synthetic 3′UTR position 3; blank for special variant/natural-target identities whose positional encoding is not explicit in the identity label.
element_position_4
Regulatory element at synthetic 3′UTR position 4; blank for special variant/natural-target identities whose positional encoding is not explicit in the identity label.
regulatory_sequence
132-nt synthetic regulatory-element sequence from Supplementary Data 1, with source capitalization retained.
regulatory_sequence_length_nt
Length of the source regulatory-element sequence in nucleotides.
library_barcode_count
Number of distinct source barcodes assigned to this reporter identity.
qc_barcode_count
Number of barcodes retained after the package read-count QC filter.
plasmid_count_rep1_mean
Mean raw plasmid-input read count per QC-passing barcode in plasmid replicate 1.
plasmid_count_rep2_mean
Mean raw plasmid-input read count per QC-passing barcode in plasmid replicate 2.
rna_count_rep1_mean
Mean raw total-RNA read count per QC-passing barcode in replicate 1.
rna_count_rep2_mean
Mean raw total-RNA read count per QC-passing barcode in replicate 2.
rna_count_rep3_mean
Mean raw total-RNA read count per QC-passing barcode in replicate 3.
rna_count_rep4_mean
Mean raw total-RNA read count per QC-passing barcode in replicate 4.
polysome_count_rep1_mean
Mean raw polysome-associated RNA read count per QC-passing barcode in replicate 1.
polysome_count_rep2_mean
Mean raw polysome-associated RNA read count per QC-passing barcode in replicate 2.
polysome_count_rep3_mean
Mean raw polysome-associated RNA read count per QC-passing barcode in replicate 3.
polysome_count_rep4_mean
Mean raw polysome-associated RNA read count per QC-passing barcode in replicate 4.
forty_s_count_rep1_mean
Mean raw 40S-associated RNA read count per QC-passing barcode in replicate 1.
forty_s_count_rep2_mean
Mean raw 40S-associated RNA read count per QC-passing barcode in replicate 2.
rna_expression_log2_rep1
Median barcode-level log2 total-RNA expression relative to the 4×blank (BBBB) control in replicate 1.
rna_expression_log2_rep2
Median barcode-level log2 total-RNA expression relative to the 4×blank (BBBB) control in replicate 2.
rna_expression_log2_rep3
Median barcode-level log2 total-RNA expression relative to the 4×blank (BBBB) control in replicate 3.
rna_expression_log2_rep4
Median barcode-level log2 total-RNA expression relative to the 4×blank (BBBB) control in replicate 4.
rna_expression_log2_median
Median of all QC-passing barcode-level log2 total-RNA expression values across the available replicates.
rna_expression_log2_mean
Arithmetic mean of all QC-passing barcode-level log2 total-RNA expression values across the available replicates.
rna_expression_log2_sd
Sample standard deviation of all QC-passing barcode-level log2 total-RNA expression values across the available replicates.
rna_expression_log2_observations
Number of barcode-by-replicate observations contributing to the total-RNA expression summary.
translation_efficiency_log2_rep1
Median barcode-level log2 translation efficiency relative to the 4×blank (BBBB) control in replicate 1.
translation_efficiency_log2_rep2
Median barcode-level log2 translation efficiency relative to the 4×blank (BBBB) control in replicate 2.
translation_efficiency_log2_rep3
Median barcode-level log2 translation efficiency relative to the 4×blank (BBBB) control in replicate 3.
translation_efficiency_log2_rep4
Median barcode-level log2 translation efficiency relative to the 4×blank (BBBB) control in replicate 4.
translation_efficiency_log2_median
Median of all QC-passing barcode-level log2 translation efficiency values across the available replicates.
translation_efficiency_log2_mean
Arithmetic mean of all QC-passing barcode-level log2 translation efficiency values across the available replicates.
translation_efficiency_log2_sd
Sample standard deviation of all QC-passing barcode-level log2 translation efficiency values across the available replicates.
translation_efficiency_log2_observations
Number of barcode-by-replicate observations contributing to the translation efficiency summary.
forty_s_association_log2_rep1
Median barcode-level log2 40S association relative to the 4×blank (BBBB) control in replicate 1.
forty_s_association_log2_rep2
Median barcode-level log2 40S association relative to the 4×blank (BBBB) control in replicate 2.
forty_s_association_log2_median
Median of all QC-passing barcode-level log2 40S association values across the available replicates.
forty_s_association_log2_mean
Arithmetic mean of all QC-passing barcode-level log2 40S association values across the available replicates.
forty_s_association_log2_sd
Sample standard deviation of all QC-passing barcode-level log2 40S association values across the available replicates.
forty_s_association_log2_observations
Number of barcode-by-replicate observations contributing to the 40S association summary.
translation_initiation_efficiency_log2_rep1
Median barcode-level log2 translation initiation efficiency (polysome/40S) relative to the 4×blank (BBBB) control in replicate 1.
translation_initiation_efficiency_log2_rep2
Median barcode-level log2 translation initiation efficiency (polysome/40S) relative to the 4×blank (BBBB) control in replicate 2.
translation_initiation_efficiency_log2_median
Median of all QC-passing barcode-level log2 translation initiation efficiency (polysome/40S) values across the available replicates.
translation_initiation_efficiency_log2_mean
Arithmetic mean of all QC-passing barcode-level log2 translation initiation efficiency (polysome/40S) values across the available replicates.
translation_initiation_efficiency_log2_sd
Sample standard deviation of all QC-passing barcode-level log2 translation initiation efficiency (polysome/40S) values across the available replicates.
translation_initiation_efficiency_log2_observations
Number of barcode-by-replicate observations contributing to the translation initiation efficiency (polysome/40S) summary.
qc_pass
TRUE for reporter identities with at least three barcodes passing the package QC filter.

Quality control

The authors retained barcode observations with >10 reads in cDNA and plasmid pools and reported high replicate correlations for total RNA, polysome, and 40S measurements. The package additionally required >10 counts in both plasmid replicates and in every available total-RNA, polysome, and 40S replicate; reporter identities needed at least three retained barcodes. All 642 identities passed the reporter-level threshold. Failed barcode observations were excluded before mean-count and log2-ratio summaries were calculated.

Curation notes

Supplementary Data 1 contains 6,500 oligonucleotide/barcode designs for 642 reporter identities; Supplementary Data 2–5 contain plasmid, total-RNA, polysome, and 40S barcode counts. The Supplementary Data 5 description says four HeLa 40S replicates, but its workbook and the Methods contain two replicates; this package uses the two deposited 40S replicates. Count-table labels Lpcx1/Lpcx2/Lpcx4 were mapped to the sequence-table labels 7pcx1/7pcx2/7pcx4. The processed table is identity-level; complete barcode-level counts remain in raw_data.

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