Experiment / E0G2N0MYMIntegrated lentiMPRA

H1 hESC lentiMPRA neural-induction time course

Identification and Massively Parallel Characterization of Regulatory Elements Driving Neural Induction

A 171-bp lentiMPRA library containing 2,664 candidate, control, and scrambled regulatory elements, with 90 designed 15-bp barcodes per element, was integrated into H1 human embryonic stem cells. Nuclear RNA and genomic DNA barcode counts were collected from three replicate cultures at 0, 3, 6, 12, 24, 48, and 72 hours after neural induction to quantify temporal reporter activity.

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

Dual-SMAD neural induction with 200 ng/mL Noggin and 10 uM SB431542; sampled at 0, 3, 6, 12, 24, 48, and 72 hours post-induction

Array-synthesized 171-bp sequences were cloned upstream of a minimal promoter and EGFP reporter in a lentiviral vector, with 90 distinct 15-bp barcodes per element split across two array halves. The library was packaged into lentivirus, infected at approximately 5-8 integrations per cell, allowed to integrate for 3 days, and assayed by sequencing barcode abundance in nuclear RNA and genomic DNA. The three reported replicates comprise two biological replicates, with one biological replicate split into two technical replicates.

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 · 83 definitions
element_id
Stable element identifier combining library class, design sequence number, and the FASTA region label
element_class
Library class: candidate, control, or scrambled negative control
design_seq_number
Sequence number used in the deposited library design
region
Region label from the plasmid-library FASTA; ordinary entries are hg19 coordinates and synthetic controls use control labels
chromosome
Chromosome or synthetic control label parsed from region
start
Start coordinate parsed from region
end
End coordinate parsed from region
length_bp
Length of the extracted regulatory sequence in base pairs
sequence_171bp
171-bp regulatory sequence extracted from the Array1 oligo design
array_sequence_discordant
Whether the Array1 and Array2 FASTA records contain different 171-bp sequences for this element
designed_barcode_count
Number of designed barcode records associated with the element
design_criterion
Table S2 criterion used to select or classify the sequence
design_additional_information
Additional sequence-selection or control information from Table S2
design_references
References supplied for the design/control entry in Table S2
closest_gene
Closest gene reported for temporal MPRA elements in Table S4
mpra_temporal_cluster
MPRA temporal activity cluster from Table S4
h3k27ac_temporal_cluster
H3K27ac temporal cluster associated with the element in Table S4
atac_temporal_cluster
ATAC-seq temporal cluster associated with the element in Table S4
rna_temporal_cluster_closest_gene
RNA-seq temporal cluster of the closest gene from Table S4
rna_temporal_cluster_correlated_gene
RNA-seq temporal cluster of the most correlated gene among the four closest genes from Table S4
published_ratio_0h
Published Table S2 MPRA RNA/DNA activity ratio at 0 hours
mean_valid_barcodes_0h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 0 hours
min_valid_barcodes_0h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 0 hours
dna_cpm_0h
Mean element-level DNA barcode count normalized to counts per million across replicates at 0 hours
rna_cpm_0h
Mean element-level RNA barcode count normalized to counts per million across replicates at 0 hours
activity_ratio_0h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 0 hours
log2_activity_0h
Base-2 logarithm of activity_ratio_0h
activity_sd_0h
Sample standard deviation of the three replicate-level normalized activity ratios at 0 hours
n_replicates_0h
Number of replicates contributing to the 0-hour summary
published_ratio_3h
Published Table S2 MPRA RNA/DNA activity ratio at 3 hours
mean_valid_barcodes_3h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 3 hours
min_valid_barcodes_3h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 3 hours
dna_cpm_3h
Mean element-level DNA barcode count normalized to counts per million across replicates at 3 hours
rna_cpm_3h
Mean element-level RNA barcode count normalized to counts per million across replicates at 3 hours
activity_ratio_3h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 3 hours
log2_activity_3h
Base-2 logarithm of activity_ratio_3h
activity_sd_3h
Sample standard deviation of the three replicate-level normalized activity ratios at 3 hours
n_replicates_3h
Number of replicates contributing to the 3-hour summary
published_ratio_6h
Published Table S2 MPRA RNA/DNA activity ratio at 6 hours
mean_valid_barcodes_6h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 6 hours
min_valid_barcodes_6h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 6 hours
dna_cpm_6h
Mean element-level DNA barcode count normalized to counts per million across replicates at 6 hours
rna_cpm_6h
Mean element-level RNA barcode count normalized to counts per million across replicates at 6 hours
activity_ratio_6h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 6 hours
log2_activity_6h
Base-2 logarithm of activity_ratio_6h
activity_sd_6h
Sample standard deviation of the three replicate-level normalized activity ratios at 6 hours
n_replicates_6h
Number of replicates contributing to the 6-hour summary
published_ratio_12h
Published Table S2 MPRA RNA/DNA activity ratio at 12 hours
mean_valid_barcodes_12h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 12 hours
min_valid_barcodes_12h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 12 hours
dna_cpm_12h
Mean element-level DNA barcode count normalized to counts per million across replicates at 12 hours
rna_cpm_12h
Mean element-level RNA barcode count normalized to counts per million across replicates at 12 hours
activity_ratio_12h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 12 hours
log2_activity_12h
Base-2 logarithm of activity_ratio_12h
activity_sd_12h
Sample standard deviation of the three replicate-level normalized activity ratios at 12 hours
n_replicates_12h
Number of replicates contributing to the 12-hour summary
published_ratio_24h
Published Table S2 MPRA RNA/DNA activity ratio at 24 hours
mean_valid_barcodes_24h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 24 hours
min_valid_barcodes_24h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 24 hours
dna_cpm_24h
Mean element-level DNA barcode count normalized to counts per million across replicates at 24 hours
rna_cpm_24h
Mean element-level RNA barcode count normalized to counts per million across replicates at 24 hours
activity_ratio_24h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 24 hours
log2_activity_24h
Base-2 logarithm of activity_ratio_24h
activity_sd_24h
Sample standard deviation of the three replicate-level normalized activity ratios at 24 hours
n_replicates_24h
Number of replicates contributing to the 24-hour summary
published_ratio_48h
Published Table S2 MPRA RNA/DNA activity ratio at 48 hours
mean_valid_barcodes_48h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 48 hours
min_valid_barcodes_48h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 48 hours
dna_cpm_48h
Mean element-level DNA barcode count normalized to counts per million across replicates at 48 hours
rna_cpm_48h
Mean element-level RNA barcode count normalized to counts per million across replicates at 48 hours
activity_ratio_48h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 48 hours
log2_activity_48h
Base-2 logarithm of activity_ratio_48h
activity_sd_48h
Sample standard deviation of the three replicate-level normalized activity ratios at 48 hours
n_replicates_48h
Number of replicates contributing to the 48-hour summary
published_ratio_72h
Published Table S2 MPRA RNA/DNA activity ratio at 72 hours
mean_valid_barcodes_72h
Mean number of shared, correctly designed RNA/DNA barcodes across the three replicates at 72 hours
min_valid_barcodes_72h
Minimum number of shared, correctly designed RNA/DNA barcodes across the three replicates at 72 hours
dna_cpm_72h
Mean element-level DNA barcode count normalized to counts per million across replicates at 72 hours
rna_cpm_72h
Mean element-level RNA barcode count normalized to counts per million across replicates at 72 hours
activity_ratio_72h
Mean replicate-level median-normalized RNA/DNA activity ratio recomputed from the deposited counts at 72 hours
log2_activity_72h
Base-2 logarithm of activity_ratio_72h
activity_sd_72h
Sample standard deviation of the three replicate-level normalized activity ratios at 72 hours
n_replicates_72h
Number of replicates contributing to the 72-hour summary

Quality control

The paper retained correctly sized 15-bp barcodes matched to the designed library and observed at least once in both RNA and DNA from the same sample. For this package, barcode counts were intersected within each of the 21 time-point/replicate pairs, aggregated by element, scaled to counts per million, stabilized with a pseudocount of 1, converted to RNA/DNA ratios, median-normalized within each sample, and averaged over the three replicates. An additional package-level filter required at least 5 shared, correctly designed barcodes for every element in every replicate and time point; all 2,664 elements passed and none were removed.

Curation notes

The paper's data availability statement names the broader GEO superseries GSE115046, while the dedicated MPRA submission is GSE115042. Ordinary Table S2 coordinates use a BED-like 0-based start and were joined to the FASTA labels after adding one to the start; the table retains the exact FASTA region label. One construct has discordant 171-bp Array1 and Array2 sequences; sequence_171bp stores Array1 and array_sequence_discordant marks it. Scrambled controls were assigned to Table S2 scram_seq_1 through scram_seq_200 in deposited FASTA order because their Table S2 labels do not contain genomic coordinates. The downloaded Table S4 contains 1,431 rows with temporal MPRA annotations, whereas the manuscript text also reports 1,547 temporal CRSs; temporal annotation fields are left blank when no Table S4 row is available.

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