Experiment / E3PFYH45QEpisomal Plasmid MPRA

Chr21q22 MPRA enhancer and allelic activity in inflammatory macrophages

A disease-associated gene desert directs macrophage inflammation through ETS2

An episomal MPRA library tested 114-nucleotide genomic sequences tiling the chr21q22 candidate-SNP region at 50-bp intervals, together with allele-specific constructs for candidate variants and control constructs. TPP macrophages from eight healthy donors were transfected; RNA was collected 24 hours later and compared with four input-DNA library replicates.

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

Inflammatory TPP differentiation: GM-CSF for 3 days, followed by GM-CSF plus TNFα, PGE2, and Pam3CSK4 for 3 days; no additional MPRA treatment

The pGL4.10M-derived episomal reporter used an RSV promoter adapted for primary macrophages. Oligos contained a 16-nt universal primer site, 114-nt variable genomic sequence, KpnI/XbaI sites, an 11-nt barcode, and a 17-nt universal primer site. Tiling constructs had six unique barcodes; each allelic construct had 30 unique barcodes. Barcode libraries were sequenced as 50-bp single-end Illumina HiSeq 2500 reads from mRNA and input DNA.

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 · 44 definitions
element_id
Source construct identifier.
construct_type
Source construct class: Enhancer or SNP.
library_subtype
Derived library class: target_tiling, target_allelic, positive_control, or negative_control.
rs_id
Source dbSNP or synthetic variant identifier; NA denotes not applicable.
element_tag
Source replicate/tiling tag (A, B, C, or NA).
allele_label
Source allele label: ref, alt, mult, or NA; nucleotide identities were not supplied in the barcode table.
coordinate_hg19
Source 114-nt construct interval on hg19, or NA:NA-NA for synthetic sequences.
sequence_114nt
114-nt variable genomic or synthetic sequence used in the construct.
source_barcode_count
Number of barcode rows for the construct in the deposited barcode-count file before the CPM filter.
qc_retained_barcode_count
Number of source barcode rows meeting the reproduced median-CPM >=1 criterion in both mRNA and DNA groups.
rna_1TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 1 TPP macrophage mRNA.
rna_2TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 2 TPP macrophage mRNA.
rna_3TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 3 TPP macrophage mRNA.
rna_4TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 4 TPP macrophage mRNA.
rna_5TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 5 TPP macrophage mRNA.
rna_6TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 6 TPP macrophage mRNA.
rna_7TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 7 TPP macrophage mRNA.
rna_8TPP_norm_count
Deposited quantile-normalized, collapsed construct count for donor 8 TPP macrophage mRNA.
dna_DNAv1_norm_count
Deposited quantile-normalized, collapsed construct count for input DNA replicate 1.
dna_DNAv2_norm_count
Deposited quantile-normalized, collapsed construct count for input DNA replicate 2.
dna_DNAv3_norm_count
Deposited quantile-normalized, collapsed construct count for input DNA replicate 3.
dna_DNAv4_norm_count
Deposited quantile-normalized, collapsed construct count for input DNA replicate 4.
dna_mean_norm_count
Arithmetic mean of the four deposited normalized input-DNA counts.
dna_median_norm_count
Median of the four deposited normalized input-DNA counts.
rna_mean_norm_count
Arithmetic mean of the eight deposited normalized mRNA counts.
activity_mean_log2_rna_dna
Mean across eight donors of log2(normalized mRNA count / mean normalized input-DNA count).
activity_median_log2_rna_dna
Median across eight donors of log2(normalized mRNA count / mean normalized input-DNA count).
activity_sd_log2_rna_dna
Sample standard deviation across eight donors of the donor-level log2 RNA/DNA activity values.
activity_1TPP_log2_rna_dna
Donor 1 log2(normalized mRNA count / mean normalized input-DNA count).
activity_2TPP_log2_rna_dna
Donor 2 log2(normalized mRNA count / mean normalized input-DNA count).
activity_3TPP_log2_rna_dna
Donor 3 log2(normalized mRNA count / mean normalized input-DNA count).
activity_4TPP_log2_rna_dna
Donor 4 log2(normalized mRNA count / mean normalized input-DNA count).
activity_5TPP_log2_rna_dna
Donor 5 log2(normalized mRNA count / mean normalized input-DNA count).
activity_6TPP_log2_rna_dna
Donor 6 log2(normalized mRNA count / mean normalized input-DNA count).
activity_7TPP_log2_rna_dna
Donor 7 log2(normalized mRNA count / mean normalized input-DNA count).
activity_8TPP_log2_rna_dna
Donor 8 log2(normalized mRNA count / mean normalized input-DNA count).
variant_pair_ref_element_id
Paired ref construct identifier for the same source rsID and element tag; blank when no ref/alt pair applies.
variant_pair_alt_element_id
Paired alt construct identifier for the same source rsID and element tag; blank when no ref/alt pair applies.
variant_effect_alt_minus_ref_mean_log2
Mean across eight donors of paired alt minus ref donor-level activity log2 values; descriptive derived contrast, not the paper's QuASAR-MPRA statistic.
variant_effect_alt_minus_ref_median_log2
Median across eight donors of paired alt minus ref donor-level activity log2 values.
variant_effect_alt_minus_ref_sd_log2
Sample standard deviation across eight donors of paired alt minus ref donor-level activity log2 values.
variant_effect_n_donors
Number of paired donor contrasts used for the derived alt-minus-ref summary.
overlaps_paper_active_window
True when the construct interval overlaps the paper-reported active enhancer focus chr21:40466236-40466677 (hg19).
qc_status
Inclusion status; every row is present in the deposited filtered normalized construct file.

Quality control

The study used FastQC v0.11.9, required a perfect library-barcode match followed by at least 10 bases of expected constant sequence, and considered a transfection successful when at least 70% of the oligonucleotide library had count >1. Barcodes with median counts per million <1 in either the mRNA or DNA samples were removed; identical constructs were collapsed and quantile normalized with normalize.quantiles from preprocessCore in R 4.1.0. The processed table retains the 382 construct rows in the deposited filtered normalized file and excludes the four raw elements absent from that file. No additional significance threshold was imposed on the derived activity values.

Curation notes

This is a mixed target library: the table preserves both the chr21q22 tiling enhancer constructs and allele-focused constructs, along with positive and negative controls. The paper reports a 442-bp active enhancer focus at chr21:40466236-40466677 (hg19), represented by the overlap flag. Source labels ref/alt/mult were retained exactly; no nucleotide or risk/non-risk mapping was inferred from the barcode table. The deposited normalized file has 382 rows, while the raw barcode file has 386 unique element IDs; four elements were excluded because they were not present in the deposited filtered construct table.

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