Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators
Cell-based assay is a powerful tool in the fields of drug discovery. One of the main targets of drug development is transcription factors. Hypoxia-inducible factors (HIFs) are transcription factors involved in cellular oxygen homeostasis and associated with disease pathophysiology. Currently, a high...
| Main Authors: | , , , , , |
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| Format: | Article |
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The Malaysian Society For Biochemistry and Molecular Biology
2023
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| Online Access: | http://psasir.upm.edu.my/id/eprint/107747/ |
| _version_ | 1848864979201032192 |
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| author | Liew, Sien-Yei Abd-Aziz, Noraini Stanbridge, Eric J Yusoff, Khatijah Shafee, Norazizah Mustafaa, Suhaili |
| author_facet | Liew, Sien-Yei Abd-Aziz, Noraini Stanbridge, Eric J Yusoff, Khatijah Shafee, Norazizah Mustafaa, Suhaili |
| author_sort | Liew, Sien-Yei |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Cell-based assay is a powerful tool in the fields of drug discovery. One of the main targets of drug development is transcription factors. Hypoxia-inducible factors (HIFs) are transcription factors involved in cellular oxygen homeostasis and associated with disease pathophysiology. Currently, a highly sensitive cell-based assay to measure HIF activity is still lacking. Using luciferase reporter constructs, we developed a highly sensitive, stable reporter cell line for monitoring the activity of HIF. Four copies of hypoxia response element (HRE) of the erythropoietin (EPO) gene, which is one of the targets of HIF, were used to drive the expression of the luciferase reporter. This stable reporter cell line gave a 500-fold increase in the luciferase signal in hypoxia compared to a normoxia condition. This robust increase in hypoxia response is crucial to ensure consistency and reproducibility of HIF assay results while reducing the overall assay cost. The establishment of this highly sensitive cell-based HIF assay may help expedite research in the discovery of HIF regulatory drugs. |
| first_indexed | 2025-11-15T13:57:25Z |
| format | Article |
| id | upm-107747 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T13:57:25Z |
| publishDate | 2023 |
| publisher | The Malaysian Society For Biochemistry and Molecular Biology |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1077472024-07-15T02:38:06Z http://psasir.upm.edu.my/id/eprint/107747/ Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators Liew, Sien-Yei Abd-Aziz, Noraini Stanbridge, Eric J Yusoff, Khatijah Shafee, Norazizah Mustafaa, Suhaili Cell-based assay is a powerful tool in the fields of drug discovery. One of the main targets of drug development is transcription factors. Hypoxia-inducible factors (HIFs) are transcription factors involved in cellular oxygen homeostasis and associated with disease pathophysiology. Currently, a highly sensitive cell-based assay to measure HIF activity is still lacking. Using luciferase reporter constructs, we developed a highly sensitive, stable reporter cell line for monitoring the activity of HIF. Four copies of hypoxia response element (HRE) of the erythropoietin (EPO) gene, which is one of the targets of HIF, were used to drive the expression of the luciferase reporter. This stable reporter cell line gave a 500-fold increase in the luciferase signal in hypoxia compared to a normoxia condition. This robust increase in hypoxia response is crucial to ensure consistency and reproducibility of HIF assay results while reducing the overall assay cost. The establishment of this highly sensitive cell-based HIF assay may help expedite research in the discovery of HIF regulatory drugs. The Malaysian Society For Biochemistry and Molecular Biology 2023-08-31 Article PeerReviewed Liew, Sien-Yei and Abd-Aziz, Noraini and Stanbridge, Eric J and Yusoff, Khatijah and Shafee, Norazizah and Mustafaa, Suhaili (2023) Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators. Malaysian Journal of Biochemistry and Molecular Biology, 26 (1). pp. 113-121. ISSN 15112616; ESSN: - https://1f169227-0328-4af2-99bc-24934f131be8.filesusr.com/ugd/66925b_510efce4c834460d99ebee0e4866924b.pdf |
| spellingShingle | Liew, Sien-Yei Abd-Aziz, Noraini Stanbridge, Eric J Yusoff, Khatijah Shafee, Norazizah Mustafaa, Suhaili Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| title | Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| title_full | Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| title_fullStr | Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| title_full_unstemmed | Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| title_short | Establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| title_sort | establishment of a highly sensitive cell-based assay for screening of hypoxia-inducible factor regulators |
| url | http://psasir.upm.edu.my/id/eprint/107747/ http://psasir.upm.edu.my/id/eprint/107747/ |