Experiment / E2ENR7VZ1Episomal Plasmid MPRA

Upstream promoter-variant MPRA in Saccharomyces cerevisiae BY4741

Systematic identification of cis-regulatory variants that cause gene expression differences in a yeast cross

The Upstream library tested natural BY/RM variants in 144-bp promoter fragments centered on variants located upstream of yeast genes. Six effective BY4741 replicate groups were assayed by matched plasmid-DNA and reporter-RNA barcode sequencing, with RM and BY allele oligos compared pairwise.

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

An episomal low-copy RCP83 reporter plasmid (Addgene 163466) carried synthetic 144-bp fragments centered on individual promoter variants and an invariant HIS3 minimal promoter between the fragment and codon-optimized yEGFP reporter. The library covered variants from approximately 72 bp upstream of the start codon toward the preceding gene, up to 1 kb. Reporter RNA and plasmid DNA barcode counts were measured in six effective replicate groups from BY4741 cultures; the two failed DNA samples were replaced with same-batch DNA as described by the authors. The published mpra R-package model produced the RM-versus-BY log2 fold changes and FDRs.

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 · 46 definitions
variant_id
Canonical sacCer3 BY-to-RM variant identifier (chromosome_position_reference_alternate).
promoter_gene
Systematic yeast gene whose promoter fragment was tested.
library
MPRA sub-library: TSS or Upstream.
design_type
Design class from the MPRA oligo design: firstTileUTR or singleVariantTiles.
test_name
Published limma test/construct label from Figure 2 source data 1.
oligo_id
Identifier of the designed oligo carrying the tested RM allele.
orientation
Genomic strand orientation used for the promoter fragment.
chromosome
sacCer3 chromosome.
position_saccer3
1-based sacCer3 coordinate of the variant.
reference_allele_by
BY/S288C allele in the designed reference sequence.
alternate_allele_rm
RM allele substituted for the tested contrast.
variant_type
SNV when both alleles are one base; otherwise indel.
variant_index_in_fragment
1-based index of this variant among variants in the TSS fragment (1 for Upstream).
variant_position_in_fragment
Variant position within the designed promoter fragment.
fragment_variant_count
Number of natural variants represented in the oligo fragment.
fragment_anchor_position_saccer3
TSS/anchor coordinate reported in the oligo design.
fragment_allele_pattern
Published oligo allele pattern; 1 denotes BY and 2 denotes RM.
log2_fold_change_rm_vs_by
Published log2 fold change for RM-allele versus BY-allele reporter activity.
rm_vs_by_fold_change
Linear fold change calculated as 2 raised to the published log2 fold change.
mean_log_expression
Published average normalized log expression for the tested oligo.
t_statistic
Published moderated t statistic.
p_value
Published nominal p-value for the allele-activity test.
fdr
Published multiple-testing-adjusted p-value.
b_statistic
Published limma B statistic.
significant_fdr_0_05
Whether the published FDR is at most 0.05.
effect_direction
Direction of the RM allele effect on reporter expression.
n_barcodes_assigned_to_test_oligo
Number of barcode rows assigned to the tested oligo in the GEO mapped-to-design matrix.
dna_count_sum_effective_replicates
Sum of plasmid-DNA barcode counts across the paper's effective replicate groups.
rna_count_sum_effective_replicates
Sum of reporter-RNA barcode counts across the paper's effective replicate groups.
log2_rna_dna_ratio
Descriptive log2 ratio of summed RNA barcode counts to summed DNA barcode counts; not the modeled allele effect.
min_dna_count_across_effective_replicates
Minimum aggregated DNA count across effective replicate groups; all retained rows are greater than zero.
min_rna_count_across_effective_replicates
Minimum aggregated RNA count across effective replicate groups; all retained rows are greater than zero.
dna_replicate_count
Number of effective DNA replicate groups summarized.
rna_replicate_count
Number of effective RNA replicate groups summarized.
supplementary_annotation_available
Whether a matching row was available in Supplementary file 3 annotations.
population_frequency
Population-frequency annotation supplied in Supplementary file 3.
maf
Minor-allele-frequency annotation supplied in Supplementary file 3.
derived_allele_frequency
Derived-allele-frequency annotation supplied in Supplementary file 3.
rm_derived
Indicator supplied by the paper for whether the RM allele is derived.
distance_from_gene_bp
Distance from the target gene reported in Supplementary file 3.
in_a_nucleosome
Indicator for overlap with a nucleosome annotation.
nucleosome_score
Nucleosome score supplied by the paper.
has_human_homolog
Indicator that the target gene has a human homolog.
has_paralog
Indicator that the target gene has a paralog.
overlapping_gene_count
Number of genes overlapped by the promoter annotation.
source_table
Source workbook and worksheet used for the published effect statistics.

Quality control

The authors retained barcode assignments whose primary oligo perfectly matched a designed oligo, excluded reads not matching known annotation barcodes, and replaced the two failed Upstream DNA samples with same-batch DNA measurements. They tested only oligos with summed barcode counts greater than zero in every DNA and RNA replicate. Package QC additionally required finite published statistics with p-values and FDRs in [0,1], removed the two Hunter control constructs, and retained 4,467 natural Upstream variant contrasts; every retained row has nonzero counts in all six effective DNA and six effective RNA replicate groups.

Curation notes

This table is one row per natural variant contrast, retaining separate promoter-gene/strand contexts when the same genomic variant was assayed in divergent promoters. The processed GEO count matrix was aggregated by oligo and summarized using the paper's six effective replicate groups, including the documented same-batch DNA replacements. Optional biological annotations are copied from Supplementary file 3 when a matching test row exists; blank annotation fields do not indicate failed MPRA QC.

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