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Updated: Jun 12, 2026
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Screening for Functional Non-coding Genetic Variants Using Electrophoretic Mobility Shift Assay (EMSA) and DNA-affinity Precipitation Assay (DAPA)
Published on: August 21, 2016
Functional characterization of polymorphisms in the human TFPI gene
Grethe Skretting1, Benedicte Stavik, Nina E Landvik
1Department of Hematology, Oslo University Hospital, Kirkeveien 166, Oslo, Norway. grethe.skretting@medisin.uio.no
Biochemical and Biophysical Research Communications
|June 4, 2010
Summary
Single nucleotide polymorphisms (SNPs) in the tissue factor pathway inhibitor (TFPI) gene may influence thrombosis risk. Specific alleles, -287C and -33C, show potential association with a reduced risk of thrombosis.
Area of Science:
- Genetics
- Hematology
- Molecular Biology
Background:
- Tissue factor pathway inhibitor (TFPI) is crucial for regulating coagulation.
- Low TFPI levels are linked to increased arterial and venous thrombosis risk.
- Single nucleotide polymorphisms (SNPs) in TFPI regulatory regions may impact plasma TFPI levels.
Purpose of the Study:
- To investigate the functional impact of TFPI gene polymorphisms on TFPI regulation.
- To explore the molecular mechanisms by which SNPs affect TFPI promoter activity.
- To assess the potential association of these polymorphisms with thrombosis risk.
Main Methods:
- Reporter gene assays were used to evaluate promoter activity with different TFPI SNPs.
- Electrophoretic mobility shift assays (EMSA) were employed to analyze DNA-protein interactions at polymorphic sites.
- Functional analysis of -287T/C, -399C/T, and -33T/C SNPs in TFPI regulatory regions.
Main Results:
- The -287C and -399T alleles significantly altered TFPI promoter activity.
- The -33T/C polymorphism also demonstrated an effect on transcriptional activity.
- EMSA indicated that all three polymorphisms potentially modify DNA-protein binding.
- A transcriptional association was observed between the -287C allele and the -33T/C polymorphism.
Conclusions:
- TFPI gene polymorphisms (-287T/C, -399C/T, -33T/C) functionally impact TFPI regulation.
- These SNPs alter TFPI promoter activity and DNA-protein interactions.
- The -287C and -33C alleles may be associated with a reduced risk of thrombosis.
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