Omics · study · 2026
Redox effector factor-1 (Ref-1) CRISPR-Engineered Cys93 and Cys99 Mutants Reveal Cooperative and Secondary Roles of Redox Signaling and PDAC Progression
Listed in NCBI GEO
APE1/Ref-1 is a multifunctional protein implicated in pancreatic ductal adenocarcinoma (PDAC) progression through its redox regulation of oncogenic transcription factors such as HIF-1α, NF-κB, and others.
Description
While Cys65 is recognized as the primary redox-active residue, the functions of other cysteines remain only partially understood. Here, we describe the first CRISPR-engineered homozygous PDAC cell lines with single (C93A, C99A) and double (C93A+C99A) point mutations in Ref-1.
These models enabled detailed analysis of the individual and combined roles of Cys93 and Cys99 in Ref-1 redox signaling and their impact on various cancer phenotypes. A single mutation at Cys93 led to decreased transcription factor activation and reduced tumor cell survival in vitro and in vivo. These effects were similar but not as profound in cells expressing C99A.
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The C93A+C99A double mutant further impaired redox signaling and significantly decreased tumor growth and metastasis in orthotopic PDAC mouse models. All of these perturbations specifically disrupted Ref-1’s redox activity without impacting its DNA repair function. Due to reduced levels of Ref-1 protein in the C93A+C99A tumor cells, add-back experiments were performed with wt-Ref-1 and C93A+C99A-Ref-1 to examine the impact on cell survival and activation of transcription factors downstream of Ref-1.
Transcriptomic analysis revealed distinct changes in redox and metabolic pathways, though these were less pronounced than those observed with the previously studied C65A mutant.
Links
Get the data
- GEO FTP directory ftp.ncbi.nlm.nih.gov/geo/series/GSE347nnn/GSE347008 ↗
download · from NCBI GEO
Where it is published
- GEO accession page ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE347008 ↗
landing page · from NCBI GEO
Documentation and papers
- PRJNA1527481 ncbi.nlm.nih.gov/bioproject/PRJNA1527481 ↗
project · from NCBI GEO
Topics
- Stated by source
- Expression profiling by high throughput sequencing · Homo sapiens
- From keywords
- Life Sciences
- Inferred from text
- Cancer 75%
Provenance · 1 source records, 8 field assertions
| Source | Key | Last seen | Raw |
|---|---|---|---|
| NCBI GEO | GSE347008 | 14 d ago | JSON v1 |
| Field | Assertion | Extractor | Evidence |
|---|---|---|---|
| access_level | source · NCBI GEO | connector:ncbi_geo@1.0.0 | |
| concepts[disease].local:disease:cancer | enrichment · NCBI GEO | keyword-concept-rules@1.0.0 | title+description (75%) |
| concepts[field].local:field:life-sciences | mapping · NCBI GEO | connector:ncbi_geo@1.0.0 | |
| concepts[method].geo_series_type:expression-profiling-by-high-throughput-sequencing | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /gdstype |
| concepts[organism].NCBITaxon:9606 | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /taxon |
| description | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /summary |
| publication_date | source · NCBI GEO | connector:ncbi_geo@1.0.0 | |
| title | source · NCBI GEO | connector:ncbi_geo@1.0.0 | /title |