Experiment / E1ZSECMFIAAV-MPRA / in vivo MPRA

AAV-MPRA 5-bp tiling mutagenesis of chamber-selective enhancers

In Vivo Dissection of Chamber-Selective Enhancers Reveals Estrogen-Related Receptor as a Regulator of Ventricular Cardiomyocyte Identity

A pooled AAV9 library tiled 29 natural 400-bp chamber-selective enhancers with 190-bp WT oligos and matched central 5-bp deletion mutants. The library was assayed in five atrial and four ventricular RNA replicates with five DNA input replicates to identify sequence elements required for chamber activity and selectivity.

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; AAV9 mutagenesis library delivered at P0 and heart chambers collected at P7

Each of 29 400-bp enhancers was represented by 80 WT tiles offset by 5 bp, with a paired oligo deleting the central 5 bp. The Methods text calls the WT oligos 190 nt, while the Figure 5 caption and deposited coordinate spans describe 180-bp oligos; the package preserves the deposited coordinates. Each oligo carried a unique 10-nt barcode. The library also contained negative ESC-derived controls and positive enhancer controls. GEO supplies deduplicated read counts for five DNA, five atrial RNA, and four ventricular RNA samples.

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 · 75 definitions
oligo_id
Exact deposited mutagenesis-library region identifier.
parent_enhancer_region
400-bp parent region for tiling enhancers or the source control interval.
design_id
Source design token or control name.
library_class
Source library class: A_enhancers, V_enhancers, AV_enhancers, enhancer positive controls, or ESC_enhancers.
source_region_id
Source genomic region used to identify the parent enhancer or control.
oligo_interval
180-bp oligo interval reported by the source.
tile_index
Source 5-bp tile index; 1–80 for the 400-bp tiling library and a source control token for controls.
tile_start_offset_bp
Approximate 0-based offset of the tile within the parent region, calculated as (tile index minus 1) × 5 when the tile index is 1–80.
oligo_type
WT wild-type tile or Mut tile with the central 5 bp deleted.
sequence_5mer
Deposited 5-bp sequence for WT rows; the source token NO for Mut rows.
wildtype_sequence_5mer
Wild-type 5-bp sequence for the row’s WT–Mut pair, when available.
deleted_5bp
5-bp sequence deleted in the paired Mut oligo; blank for WT/control rows.
pair_id
Stable identifier for a WT–Mut oligo pair; control-only rows receive a stable singleton identifier.
pair_complete
TRUE when both WT and Mut members passed the DNA-coverage filter.
chromosome
Chromosome parsed from the parent enhancer/control coordinate.
enhancer_start
Parent region start coordinate.
enhancer_end
Parent region end coordinate.
enhancer_length_bp
Parent coordinate span calculated as end minus start.
dna_count_rep1
Deposited DNA read count for replicate 1.
dna_count_rep2
Deposited DNA read count for replicate 2.
dna_count_rep3
Deposited DNA read count for replicate 3.
dna_count_rep4
Deposited DNA read count for replicate 4.
dna_count_rep5
Deposited DNA read count for replicate 5.
atrium_rna_count_rep1
Deposited atrial RNA read count for replicate 1.
atrium_rna_count_rep2
Deposited atrial RNA read count for replicate 2.
atrium_rna_count_rep3
Deposited atrial RNA read count for replicate 3.
atrium_rna_count_rep4
Deposited atrial RNA read count for replicate 4.
atrium_rna_count_rep5
Deposited atrial RNA read count for replicate 5.
ventricle_rna_count_rep1
Deposited ventricular RNA read count for replicate 1.
ventricle_rna_count_rep2
Deposited ventricular RNA read count for replicate 2.
ventricle_rna_count_rep3
Deposited ventricular RNA read count for replicate 3.
ventricle_rna_count_rep4
Deposited ventricular RNA read count for replicate 4.
dna_fpm_rep1
DNA replicate 1 read count normalized to fragments per million within that sample.
dna_fpm_rep2
DNA replicate 2 read count normalized to fragments per million within that sample.
dna_fpm_rep3
DNA replicate 3 read count normalized to fragments per million within that sample.
dna_fpm_rep4
DNA replicate 4 read count normalized to fragments per million within that sample.
dna_fpm_rep5
DNA replicate 5 read count normalized to fragments per million within that sample.
atrium_rna_fpm_rep1
atrial RNA replicate 1 read count normalized to fragments per million within that sample.
atrium_rna_fpm_rep2
atrial RNA replicate 2 read count normalized to fragments per million within that sample.
atrium_rna_fpm_rep3
atrial RNA replicate 3 read count normalized to fragments per million within that sample.
atrium_rna_fpm_rep4
atrial RNA replicate 4 read count normalized to fragments per million within that sample.
atrium_rna_fpm_rep5
atrial RNA replicate 5 read count normalized to fragments per million within that sample.
ventricle_rna_fpm_rep1
ventricular RNA replicate 1 read count normalized to fragments per million within that sample.
ventricle_rna_fpm_rep2
ventricular RNA replicate 2 read count normalized to fragments per million within that sample.
ventricle_rna_fpm_rep3
ventricular RNA replicate 3 read count normalized to fragments per million within that sample.
ventricle_rna_fpm_rep4
ventricular RNA replicate 4 read count normalized to fragments per million within that sample.
dna_fpm_mean
Mean DNA fragments per million across five DNA replicates.
dna_fpm_max
Maximum DNA fragments per million across five DNA replicates; individual-oligo QC requires ≥5.
atrium_rna_fpm_mean
Mean atrial RNA fragments per million across five replicates.
ventricle_rna_fpm_mean
Mean ventricular RNA fragments per million across four replicates.
atrium_activity_log2
Mean per-replicate log2((atrial RNA FPM + 0.5)/(DNA FPM + 0.5)).
ventricle_activity_log2
Mean per-replicate log2((ventricular RNA FPM + 0.5)/(DNA FPM + 0.5)).
atrium_vs_ventricle_log2
Atrial activity log2 score minus ventricular activity log2 score.
atrium_vs_ventricle_ratio
Two raised to atrium_vs_ventricle_log2; values above 1 favor atrial activity.
atrium_activity_p_value
Two-sided one-sample t-test p-value for atrial activity scores versus zero.
ventricle_activity_p_value
Two-sided one-sample t-test p-value for ventricular activity scores versus zero.
atrium_active_nominal
Nominal activity flag using a positive score and unadjusted p<0.05; not the paper’s motif-call criterion.
ventricle_active_nominal
Nominal activity flag using a positive score and unadjusted p<0.05; not the paper’s motif-call criterion.
paired_wt_activity_atrium_log2
Atrial activity score of the WT member of the complete pair.
paired_mut_activity_atrium_log2
Atrial activity score of the Mut member of the complete pair.
paired_mut_minus_wt_atrium_log2
Mut minus WT atrial activity log2 score.
paired_mut_minus_wt_atrium_p_value
Two-sided one-sample t-test p-value for per-replicate Mut minus WT atrial activity differences.
paired_mut_minus_wt_atrium_fdr
Benjamini-Hochberg FDR across complete-pair atrial effects.
paired_wt_activity_ventricle_log2
Ventricular activity score of the WT member of the complete pair.
paired_mut_activity_ventricle_log2
Ventricular activity score of the Mut member of the complete pair.
paired_mut_minus_wt_ventricle_log2
Mut minus WT ventricular activity log2 score.
paired_mut_minus_wt_ventricle_p_value
Two-sided one-sample t-test p-value for per-replicate Mut minus WT ventricular activity differences.
paired_mut_minus_wt_ventricle_fdr
Benjamini-Hochberg FDR across complete-pair ventricular effects.
paired_wt_selectivity_log2
WT atrial-minus-ventricular activity log2 score.
paired_mut_selectivity_log2
Mut atrial-minus-ventricular activity log2 score.
paired_mut_minus_wt_selectivity_log2
Change in chamber selectivity caused by the 5-bp deletion: Mut selectivity minus WT selectivity.
paired_atrium_effect_significant_fdr_0_05
TRUE when the paired atrial deletion effect has FDR<0.05.
paired_ventricle_effect_significant_fdr_0_05
TRUE when the paired ventricular deletion effect has FDR<0.05.
qc_individual_dna_pass
TRUE when at least one DNA replicate has ≥5 FPM before pair-completeness filtering.
qc_pass
TRUE for every retained row; low-coverage oligos and unpaired low-coverage Mut rows are excluded.

Quality control

The article explicitly excludes oligos with <5 FPM DNA coverage. The package applies the matching max-across-five-DNA-replicates ≥5 FPM gate, retains all passing WT oligos, and retains a Mut oligo only when its matching WT also passes, so the complete paired set is directly interpretable. This yields 2,394 WT rows and 1,761 complete WT–Mut pairs, matching the paper; rows are excluded before table.csv generation if they fail the individual or pair-completeness gate. Paired deletion effects are calculated from replicate-level log2 RNA/DNA differences and BH-adjusted across complete pairs.

Curation notes

AAV9 libraries were delivered systemically to P0 mouse pups and atria and ventricles were collected at P7. The combined biosample is represented as UNMAPPED because the same library was read out in two distinct primary cardiomyocyte populations; the constituent Cell Ontology terms are CL:0002129 (regular atrial cardiac myocyte) and CL:0002131 (regular ventricular cardiac myocyte). The deposited table contains 5,066 oligo rows, including 29 tiling enhancers plus ESC negative controls and positive enhancer controls. After the coverage and pair-completeness filters, table.csv contains 4,155 rows: 2,394 WT rows and one Mut row for each of 1,761 complete pairs. There is a source-length discrepancy: the Methods text says 190-nt oligos, but the Figure 5 caption and GEO coordinate spans use 180 bp; this package retains the GEO coordinates and does not infer unreported sequence bases. Mutagenesis count tables do not include the full WT or mutant sequence; only the WT 5-mer and deleted 5-bp token are available, so full mutant sequences are not inferred. The article’s motif-enrichment analysis is not recomputed; pair-level activity effects are provided as a reusable quantitative summary.

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