Experiment / E1Y68DEF9Deep Mutational Scanning MPRA (DMS-MPRA)

IRF6 enhancer saturation-mutagenesis MPRA in HaCaT

Integration of multiple epigenomic marks improves prediction of variant impact in saturation mutagenesis reporter assay

A saturation-mutagenized 600-bp human IRF6 enhancer library was assayed in HaCaT keratinocytes with three independent plasmid transfections. Variant reporter activity was estimated from matched RNA and DNA tag counts.

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

The episomal pGL4-derived enhancer reporter carried the error-prone-PCR-mutagenized IRF6 sequence upstream of a minimal promoter and a random sequence tag in the reporter 3′ UTR. The library was transfected into HaCaT for 24 h; matched DNA/RNA tag counts from three independent transfections were fit with log2(RNA) ~ log2(DNA) + N + offset to estimate each variant coefficient.

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 · 16 definitions
element_id
Paper Table 1 regulatory element name.
source_element_id
Element label in the authors' MPRA saturation-mutagenesis portal.
variant_id
Stable package identifier combining element, GRCh38 coordinate, reference allele, and alternate allele.
chromosome
GRCh38 chromosome with chr prefix.
position_grch38
1-based GRCh38 position of the substituted nucleotide.
position_in_element
1-based position within the paper's reported target interval.
reference_allele
Reference/template nucleotide in the tested library.
alternate_allele
Single-nucleotide substitution introduced by saturation mutagenesis.
variant_class
Variant class; all retained rows are SNV.
barcode_count
Number of unique random tags associated with the variant.
dna_count
Source count of DNA sequences carrying the variant.
rna_count
Source count of RNA sequences carrying the variant.
log2_variant_effect
Portal-fitted regression coefficient estimating the log2 RNA/DNA reporter effect of the variant.
p_value
P-value for the fitted variant coefficient.
effect_direction
Direction derived from the coefficient sign: up, down, or no_change.
significant_p_lt_1e-5
Boolean indicating whether p_value is below the paper's nominal 1e-5 threshold.

Quality control

The source saturation-mutagenesis analysis used a minimum of 10 associated tags per variant. Package QC retained 1,676 of 1,783 candidate SNVs in the paper's Table 1 interval: GRCh38 SNVs with barcode_count >= 10, positive DNA/RNA counts, and finite coefficient and p-value were retained; 107 low-tag rows plus one-base deletions and out-of-interval records were excluded. The paper/CAGI analysis used p < 1e-5 as the nominal significance threshold, exposed as significant_p_lt_1e-5.

Curation notes

HaCaT is Cellosaurus CVCL:0038. The source portal contains a 601-bp flanking interval for IRF6; table.csv is restricted to the 600-bp interval reported by the paper.

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