Experiment / E2CAWGDW35' UTR / Translation Efficiency MPRA (MPTA)

Fixed-end N50 5′UTR polysome-profiling MPRA in primary human T cells

Optimizing 5’UTRs for mRNA-delivered gene editing using deep learning

The fixed-end N50 IVT EGFP reporter library was transfected into activated human T cells enriched from healthy-donor PBMCs. Two biological replicates were fractionated by polysome load and summarized as sequence-level MRL values from UMI counts.

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

Anti-CD3/CD28 activation with IL-2

The library was unmodified IVT mRNA with a constant 25-nt 5′ prefix, fully randomized 50-nt 5′UTR segment, EGFP CDS, and bovine growth hormone-derived 3′ UTR. PBMC-derived T cells were activated with anti-CD3/CD28 and IL-2, transfected with the reporter library, treated with cycloheximide for lysis, separated by sucrose-gradient polysome profiling, and sequenced using fraction-specific barcodes and UMIs. MRL is the author-provided translation-efficiency proxy calculated from normalized fraction counts.

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 · 13 definitions
element_id
Stable package identifier assigned in lexicographic UTR-sequence order after QC.
utr_sequence
The variable 50-nt 5′UTR sequence represented by the reporter library.
utr_length
Length of the packaged variable UTR sequence in nucleotides.
replicate_count
Number of biological replicates with a finite MRL value for the sequence.
total_umi_rep1
Sum of processed UMI counts across all polysome fractions in T-cell replicate 1.
mrl_rep1
Author-calculated mean ribosome load for T-cell replicate 1.
total_umi_rep2
Sum of processed UMI counts across all polysome fractions in T-cell replicate 2.
mrl_rep2
Author-calculated mean ribosome load for T-cell replicate 2.
mrl_mean
Arithmetic mean of the replicate MRL values.
mrl_sd
Sample standard deviation of replicate MRL values.
mrl_weighted_mean
Total-UMI-weighted mean of replicate MRL values.
total_umi_sum
Sum of total UMI counts across the packaged biological replicates.
total_umi_min
Minimum total UMI count across the packaged biological replicates.

Quality control

The package follows the authors’ fixed-end comparison workflow: reported UTR strings were truncated to the first 50 nt, and only sequences with total read count >100 in all five fixed-end datasets (two HEK293T replicates, two T-cell replicates, and one HepG2 replicate) were retained. The package additionally required a 50-nt A/C/G/T sequence and finite source MRL values, leaving 204,803 shared UTRs.

Curation notes

The two T-cell processed files are GSE232927 replicate 1 and replicate 2. The article Methods describes lysis 8 h after transfection; the GEO sample text states 12 h for T-cell lysis, so this timing is retained as an unresolved source annotation rather than silently harmonized. Human T cell was resolved to Cell Ontology CL:0000084.

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