Experiment / E8N7WJEGTAAV-MPRA / in vivo MPRA

AAV-MPRA screen of chamber-selective enhancer candidates

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

A pooled AAV9 MPRA library of 400-bp mouse candidate regulatory regions was delivered to P0 pups and assayed in purified neonatal atrial and ventricular cardiomyocytes at P7. Five DNA, five atrial RNA, and five ventricular RNA replicates quantify enhancer activity and chamber 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 library delivered at P0 and heart chambers collected at P7

The AAV-MPRA uses a STARR-seq-style reporter with each candidate enhancer in the 3′ UTR downstream of an hsp68 minimal promoter and mCherry. Each 400-bp region was synthesized as a 230-nt self-priming oligonucleotide pair. Atria and ventricles were analyzed separately from the same library; GEO supplies deduplicated read counts for each region and sample.

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 · 56 definitions
region_id
Exact GEO region identifier containing the occupancy design token and mm10 coordinate.
chromosome
Chromosome parsed from the reported mm10 coordinate.
start
Reported region start coordinate; the source uses a 400-bp span where end minus start equals 400.
end
Reported region end coordinate; the source uses a 400-bp span where end minus start equals 400.
element_length_bp
Coordinate span calculated as end minus start.
occupancy_design
Source design/occupancy token before the coordinate, encoding chamber-specific or shared TF/P300 features.
library_group
GEO library category: TF1-2, TF3-4, MTF or P300, spMTF or spP300, or Neg.
dna_count_rep1
Deposited AAV-genome DNA read count for DNA replicate 1.
dna_count_rep2
Deposited AAV-genome DNA read count for DNA replicate 2.
dna_count_rep3
Deposited AAV-genome DNA read count for DNA replicate 3.
dna_count_rep4
Deposited AAV-genome DNA read count for DNA replicate 4.
dna_count_rep5
Deposited AAV-genome DNA read count for DNA replicate 5.
atrium_rna_count_rep1
Deposited atrial RNA read count for biological replicate 1.
atrium_rna_count_rep2
Deposited atrial RNA read count for biological replicate 2.
atrium_rna_count_rep3
Deposited atrial RNA read count for biological replicate 3.
atrium_rna_count_rep4
Deposited atrial RNA read count for biological replicate 4.
atrium_rna_count_rep5
Deposited atrial RNA read count for biological replicate 5.
ventricle_rna_count_rep1
Deposited ventricular RNA read count for biological replicate 1.
ventricle_rna_count_rep2
Deposited ventricular RNA read count for biological replicate 2.
ventricle_rna_count_rep3
Deposited ventricular RNA read count for biological replicate 3.
ventricle_rna_count_rep4
Deposited ventricular RNA read count for biological replicate 4.
ventricle_rna_count_rep5
Deposited ventricular RNA read count for biological replicate 5.
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.
ventricle_rna_fpm_rep5
ventricular RNA replicate 5 read count normalized to fragments per million within that sample.
dna_fpm_mean
Mean DNA fragments per million across the five DNA replicates.
dna_fpm_max
Maximum DNA fragments per million across the five DNA replicates; the package QC gate is ≥5.
atrium_rna_fpm_mean
Mean atrial RNA fragments per million across five RNA replicates.
ventricle_rna_fpm_mean
Mean ventricular RNA fragments per million across five RNA 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 per-replicate log2 RNA/DNA scores versus zero.
atrium_activity_fdr
Benjamini-Hochberg FDR for the atrial activity p-values among QC-passed regions.
ventricle_activity_p_value
Two-sided one-sample t-test p-value for ventricular per-replicate log2 RNA/DNA scores versus zero.
ventricle_activity_fdr
Benjamini-Hochberg FDR for the ventricular activity p-values among QC-passed regions.
chamber_selectivity_p_value
Welch two-sample p-value comparing atrial and ventricular per-replicate log2 RNA/DNA scores.
chamber_selectivity_fdr
Benjamini-Hochberg FDR for the chamber-selectivity p-values among QC-passed regions.
active_atrium
Derived atrial activity call: positive activity score and FDR <0.05.
active_ventricle
Derived ventricular activity call: positive activity score and FDR <0.05.
active_any
TRUE when either chamber has a derived active call.
chamber_selective_class
Derived activity class using active calls, |atrial minus ventricular activity| >0.58, and selectivity FDR <0.05.
qc_pass
TRUE for every row retained after the source DNA-coverage filter.

Quality control

The article states that candidate regions with low AAV-genome DNA coverage were removed. Normalizing each of the five DNA libraries to FPM and retaining a region when at least one DNA replicate was ≥5 FPM reproduces the paper’s reported surviving set exactly: 2,160 candidate regions plus 479 negative-control regions (2,639 total from 3,820 deposited rows). Activity scores are mean per-replicate log2((RNA FPM + 0.5)/(DNA FPM + 0.5)); derived activity and chamber-selectivity p-values use two-sided t-tests and are BH-adjusted. Rows failing the DNA gate are absent from table.csv.

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 GEO aggregate has 3,820 rows and five library groups; this differs from the broader design totals described in the article, but the reported post-coverage counts are recovered exactly. GEO count tables do not include the full 400-bp sequences, so region coordinates and source occupancy tokens are retained while sequence strings are not fabricated. The article’s own active/CSE calls use its custom MPRA analysis; the p-values/FDR and classes in this package are transparent replicate-level summaries of the deposited counts.

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