Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities

Hypoxia-inducible factor (HIF) is one of the most important transcription factors involves in cells survival under low oxygen concentrations, termed hypoxia. Hypoxia is associated with various human diseases from ischemic injuries to cancer. The central role of HIF in these conditions has made it a...

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Main Author:	Liew, Sien Yei
Format:	Thesis
Language:	English
Published:	2013
Subjects:	Anoxemia Luciferases Erythropoietin
Online Access:	http://psasir.upm.edu.my/id/eprint/39929/1/FBSB%202013%2020R.pdf
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id	my-upm-ir.39929
record_format	uketd_dc
spelling	my-upm-ir.399292016-01-21T07:28:13Z Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities 2013-11 Liew, Sien Yei Hypoxia-inducible factor (HIF) is one of the most important transcription factors involves in cells survival under low oxygen concentrations, termed hypoxia. Hypoxia is associated with various human diseases from ischemic injuries to cancer. The central role of HIF in these conditions has made it as one of the main target molecules for drug discovery. Demand for HIF assay system is high. Cell-based HIF assay systems are preferred since they can provide direct data for intracellular biochemical changes. HIF assay systems which are currently available in the market offer limited sensitivity with restricted reproducibility at a high cost. Therefore, in the present study, a HIF assay was succesfully developed. The system used a Saos-2 human osteosarcoma cell line which is one of the most studied cell lines to express a luciferase protein in response to HIF activation. The luciferase gene construct which was previously cloned downstream of four copies of a hypoxia response element (HRE) of the erythropoietin (EPO) gene. EPO is one of the specific target genes of HIF. Therefore this assay can be used to measure HIF activation upon induction by candidate drugs of interest, by measuring the luciferase expression. This highly sensitive assay system offers a 40-times increased sensitivity compared to the currently available systems. This aspect would be highly beneficial to researchers since the increase in sensitivity can contribute towards result consistency and reproducibility leading to reduced wastage of precious drug candidates. Anoxemia Luciferases Erythropoietin 2013-11 Thesis http://psasir.upm.edu.my/id/eprint/39929/ http://psasir.upm.edu.my/id/eprint/39929/1/FBSB%202013%2020R.pdf application/pdf en public masters Universiti Putra Malaysia Anoxemia Luciferases Erythropoietin
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Anoxemia Luciferases Erythropoietin
spellingShingle	Anoxemia Luciferases Erythropoietin Liew, Sien Yei Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
description	Hypoxia-inducible factor (HIF) is one of the most important transcription factors involves in cells survival under low oxygen concentrations, termed hypoxia. Hypoxia is associated with various human diseases from ischemic injuries to cancer. The central role of HIF in these conditions has made it as one of the main target molecules for drug discovery. Demand for HIF assay system is high. Cell-based HIF assay systems are preferred since they can provide direct data for intracellular biochemical changes. HIF assay systems which are currently available in the market offer limited sensitivity with restricted reproducibility at a high cost. Therefore, in the present study, a HIF assay was succesfully developed. The system used a Saos-2 human osteosarcoma cell line which is one of the most studied cell lines to express a luciferase protein in response to HIF activation. The luciferase gene construct which was previously cloned downstream of four copies of a hypoxia response element (HRE) of the erythropoietin (EPO) gene. EPO is one of the specific target genes of HIF. Therefore this assay can be used to measure HIF activation upon induction by candidate drugs of interest, by measuring the luciferase expression. This highly sensitive assay system offers a 40-times increased sensitivity compared to the currently available systems. This aspect would be highly beneficial to researchers since the increase in sensitivity can contribute towards result consistency and reproducibility leading to reduced wastage of precious drug candidates.
format	Thesis
qualification_level	Master's degree
author	Liew, Sien Yei
author_facet	Liew, Sien Yei
author_sort	Liew, Sien Yei
title	Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
title_short	Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
title_full	Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
title_fullStr	Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
title_full_unstemmed	Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
title_sort	establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities
granting_institution	Universiti Putra Malaysia
publishDate	2013
url	http://psasir.upm.edu.my/id/eprint/39929/1/FBSB%202013%2020R.pdf
_version_	1747811826487263232

Establishment of a cell-based reporter assay for screening of hypoxia-inducible factor activities

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