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Journal: Human Genetics and Genomics Advances
Article Title: A non-coding ABO regulatory variant associatedwith VWF levels, thrombosis risk, and COVID-19 severity is topologically linked to ADAMTS13 in endothelial cells
doi: 10.1016/j.xhgg.2025.100550
Figure Lengend Snippet: Chromatin interactions and regulatory landscape at the ABO and ADAMTS13 loci in endothelial cells (A) Hi-C data from HUVECs showing chromatin interactions within a segment of chromosome 9, encompassing the ABO and ADAMTS13 loci. Three loop boundaries (L1, L2, L3) are identified, indicating regions anchoring chromatin loops. (B) Detailed epigenomic landscape of the ABO and ADAMTS13 loci, corresponding to the region amplified in (C). Data from multiple assays are shown. RNA-seq data from HUVEC showing gene expression levels at the ABO and ADAMTS13 loci. ADAMTS13 is expressed in HUVECs, whereas ABO shows no detectable expression in this cell type. ChIA-PET (CTCF): identifies CTCF-binding at L1, L2, and L3, suggesting these sites demarcate loop boundaries. DNase-seq: reveals accessible chromatin regions aligning with the loop anchors. ChIP-seq for RAD21 (cohesin): confirms cohesin enrichment at L1, L2, and L3. ChIP-seq data for HBMECs identify CTCF binding sites, while DNase-seq reveals accessible chromatin regions at L1, L2, and L3. (C) Drawn chromatin interaction model of the ABO and ADAMTS13 loci based on epigenomic and chromatin conformation data integrating Hi-C, ChIA-PET, DNase-seq, and ChIP-seq data from HUVEC, created with BioRender.com .
Article Snippet: Quantitative PCR was conducted with TaqMan Fast Advanced Master Mix (Applied Biosystems, catalog no. 4444556) and TaqMan probes for ADAMTS13 (Applied Biosystems, catalog no.
Techniques: Hi-C, Amplification, RNA Sequencing, Gene Expression, Expressing, ChIA Pet Assay, Binding Assay, ChIP-sequencing
Journal: Human Genetics and Genomics Advances
Article Title: A non-coding ABO regulatory variant associatedwith VWF levels, thrombosis risk, and COVID-19 severity is topologically linked to ADAMTS13 in endothelial cells
doi: 10.1016/j.xhgg.2025.100550
Figure Lengend Snippet: Functional testing and motif evaluation of non-coding variants at the ABO locus (A) Tables summarizing phenotypes previously associated with the non-coding variants rs657152 and rs505922 at the ABO locus. (B) Results of CRISPRa activation assays targeting the regions containing rs657152 and rs505922 in HUVECs. Three sgRNAs were used to activate transcription at each locus. The region harboring rs657152 demonstrated a significant increase in ADAMTS13 expression ( p < 0.05), supporting its role as a cis -regulatory element. In contrast, targeting the rs505922 region showed no significant activation. Data are represented as average fold change relative to control, with each point being a technical replicate ( n = 3). (C) Results of luciferase reporter assays for the genomic regions containing rs657152 and rs505922. For each variant, constructs containing either the risk or non-risk allele were tested in HUVECs to evaluate allele-specific transcriptional activity. The assay showed that the risk allele of rs505922 significantly reduced luciferase activity compared to the non-risk allele ( p < 0.05). In contrast, no statistically significant difference was observed between the alleles of rs657152. Data are represented as average luciferase activity relative to control, normalized with Renilla luciferase activity, with each point being a technical replicate ( n = 3) for each variant. (D) Motif analysis for the genomic regions containing rs657152 and rs505922. The in silico analysis demonstrated allele-specific transcription factor binding motifs for both variants. Substitution of the risk and non-risk alleles resulted in distinct motif profiles.
Article Snippet: Quantitative PCR was conducted with TaqMan Fast Advanced Master Mix (Applied Biosystems, catalog no. 4444556) and TaqMan probes for ADAMTS13 (Applied Biosystems, catalog no.
Techniques: Functional Assay, Activation Assay, Expressing, Control, Luciferase, Variant Assay, Construct, Activity Assay, In Silico, Binding Assay