Experiment / E5U1Y1QN9Other

Dual-luciferase validation of six STARR-seq candidate enhancers

Influence of genetic polymorphism on transcriptional enhancer activity in the malaria vector Anopheles coluzzii

Each candidate interval was cloned into an episomal pGL-Gateway-DSCP firefly-luciferase reporter and co-transfected with pRL-ubi-63E Renilla control in A. coluzzii 4a3A cells. Activity was normalized to the DLX negative-control construct.

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Perturbation & assay details

Basal / Untreated; 24 h after reporter transfection

96-well assay with 1 × 10^5 4a3A cells per well; six-fold replication within a plate across at least two independent plates; values were corrected by Renilla and normalized to the same-plate DLX mean.

Episomal dual-luciferase reporter validation: candidate insert in pGL-Gateway-DSCP drives firefly luciferase, pRL-ubi-63E provides a Renilla internal control, and the readout is normalized firefly/Renilla RLU rather than barcode counts; this is not a barcode-count MPRA.

Processed data

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Visible columns (17 of 17)
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Column dictionary · 17 definitions
element_id
Paper element identifier.
proximal_gene
Nearest annotated gene used to name the candidate enhancer.
gene_id
Gene identifier reported in Table 1.
chromosome
Chromosome arm in PEST AgamP4 coordinates.
coordinate_start
1-based inclusive interval start from Table 1.
coordinate_end
1-based inclusive interval end from Table 1.
interval_length_bp
Inclusive interval length calculated from the published coordinates.
construct_label
Candidate construct tested in the luciferase validation panel.
reporter_vector
Firefly reporter and Renilla internal-control plasmids.
cell_line
Reporter-assay cell line and Cellosaurus identifier.
activity_measure
Paper’s within-well Renilla-corrected firefly activity normalized to DLX background.
relative_luciferase_activity_median_approx
Approximate median relative luciferase activity digitized from the Figure 2 median bar; not a machine-readable source value.
n_measurements
Plotted well measurements; methods report six-fold replication across at least two plates, yielding at least 12 measurements.
p_value_vs_DLX
Paper-reported significance threshold for each candidate versus DLX background.
above_DLX_background
Paper’s candidate-level above-background call.
source_figure
Publication figure supporting the measurement.
qc_pass
Retained because the paper reports p < 0.005 versus DLX for all six candidates and at least 12 wells per condition.

Quality control

Applied the paper’s QC: within-well Renilla correction, DLX background normalization, at least 12 measurements per candidate, and non-parametric comparison to DLX. All six candidate constructs passed the reported p < 0.005 threshold.

Curation notes

The paper publishes these values only as plotted points/median bars. Values in the activity column are approximate digitizations from the publisher Figure 2 PNG retained in raw_data; they should not be treated as exact raw RLU values.

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