A Luciferase Reporter Assay to Detect Cellular Hypoxia In Vitro.

Masayuki Inubushi, Yasuto Takeuchi, Chika Murai, Yoshimasa Kitagawa

Abstract read

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Article in Methods in molecular biology (Clifton, N.J.), 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

4 authors.

Masayuki InubushiDivision of Nuclear Medicine, Department of Radiology, Kawasaki Medical School, Kurashiki, Okayama, Japan. inubushi@med.kawasaki-m.ac.jp.

Yasuto TakeuchiDivision of Cancer Cell Biology, Innovative Cancer Model Research Center, Cancer Research Institute of Kanazawa University, Kanazawa, Ishikawa, Japan.

Chika MuraiOral Diagnosis and Oral Medicine, Graduate School of Dental Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.

Yoshimasa KitagawaOral Diagnosis and Oral Medicine, Graduate School of Dental Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Hypoxia is a hallmark of ischemic cardiovascular diseases and solid malignant tumors. Cellular hypoxia induces numerous physiological and pathological processes, including hematopoiesis, angiogenesis, metabolic changes, cell growth, and apoptosis. Hypoxia-inducible factor-1 (HIF-1) binds to hypoxia response elements (HREs) to selectively induce the expression of various genes in response to hypoxia. Therefore, HREs have been used to develop hypoxia-targeted gene therapy.More than 70 pairs of HREs and hypoxia-inducible genes have been identified. The hypoxia-induced gene expression levels vary among HRE sequences depending on the number of HRE copies and oxygen levels. Most known HREs have not yet been thoroughly studied. Recent studies have revealed that the HRE-mediated effects of hypoxia are cell line-dependent. Herein we describe an in vitro method to investigate gene activation levels and characteristics based on varying the copy number of HREs in response to cellular hypoxia. We explain how to clone HREs into luciferase reporter constructs in the sense, antisense, and dual directions to measure luciferase expression for functional analyses.

Indexed as

HypoxiaOxygenApoptosisCell HypoxiaHumansLuciferasesLuciferasesOxygenBioluminescenceCell cultureCellular hypoxiaFirefly luciferaseHypoxia response element (HRE)Molecular imagingRenilla luciferase

Identifiers

PMID38319570

DOI10.1007/s12149-014-0901-2

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.