Experiment / E9U78YZ9ATargeted Genomic Integration MPRA

High-CpG E. coli fragment library with housekeeping controls

CG dinucleotides enhance promoter activity independent of DNA methylation

TrAC-seq measured wild-type activity for E. coli genomic fragments selected for high CpG density, mixed with mouse housekeeping-promoter controls, in TC-1 mouse embryonic stem cells. The assay tests whether CpG density alone is sufficient to drive transcription in chromatin.

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Perturbation & assay details

Basal / untreated; wild-type TC-1 mouse embryonic stem cells

TrAC-seq used pooled promoter-barcode cassettes inserted by recombinase-mediated cassette exchange into the beta-globin locus. RNA/cDNA and genomic-DNA barcodes were sequenced separately; per-barcode RNA/DNA enrichment used the study pseudocount alpha=0.05 and was summarized across barcodes and three biological replicates. The library contained a CpG-free eGFP spacer and a unique barcode.

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 · 15 definitions
element_id
Identifier for an E. coli genomic fragment (source interval/orientation label) or a mixed-library housekeeping-promoter control.
library
Source expression library name.
gc_percent
GC content of the tested fragment, in percent.
cpg_percent
CpG content of the tested fragment, as reported by the source.
cpg_observed_expected
Observed-over-expected CpG density (OE) for the tested fragment.
activity_rep1
RNA/DNA barcode enrichment for biological replicate 1; source activity includes the study pseudocount.
dna_barcodes_rep1
Number of DNA-supported barcodes contributing to replicate 1 activity.
activity_rep2
RNA/DNA barcode enrichment for biological replicate 2; source activity includes the study pseudocount.
dna_barcodes_rep2
Number of DNA-supported barcodes contributing to replicate 2 activity.
activity_rep3
RNA/DNA barcode enrichment for biological replicate 3; source activity includes the study pseudocount.
dna_barcodes_rep3
Number of DNA-supported barcodes contributing to replicate 3 activity.
activity_geomean
Geometric mean of the non-missing per-replicate activity enrichments.
log2_activity
Mean log2-transformed activity enrichment across non-missing replicates.
mean_dna_barcodes
Arithmetic mean of numeric DNA-supported barcode counts across replicates.
qc_replicates
Number of replicates with activity and at least 3 DNA-supported barcodes; retained rows have at least 2.

Quality control

The paper retained uniquely assigned promoter barcodes (or assignments with second-most abundant sequence / most abundant sequence <0.3), required >20 normalized genomic-DNA reads per barcode, assigned absent RNA observations zero counts, and used alpha=0.05. For this packaged element table, rows were retained only when activity and at least 3 DNA-supported barcodes were available in at least 2 of 3 biological replicates; 17 of 83 source rows passed this element-level filter.

Curation notes

Input: raw_data/GSE116704_CpGDensitylib1_results.tab.gz; GEO reporter samples GSM3258747-GSM3258753. The library mixes non-mouse E. coli genomic fragments with mouse housekeeping controls, so mm9 is recorded for the mouse assay context and E. coli entries have no mouse coordinates. The source file contains 83 rows; 17 passed the packaged QC.

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