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A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Published on: July 3, 2013
Hypoxia activates constitutive luciferase reporter constructs
Diane M Doran1, Kashmira Kulkarni-Datar, David R Cool
1Program in Microbiology and Immunology, Wright State University, Boonshoft School of Medicine, Dayton, OH 45435, USA.
Biochimie
|October 26, 2010
Summary
Hypoxia significantly increases reporter gene activity in placental cells, impacting studies on oxygen regulation. This finding suggests caution when interpreting transcriptional activity in hypoxic conditions using dual-reporter assays.
Area of Science:
- Cell Biology
- Developmental Biology
- Physiology
Background:
- Hypoxia, or low oxygen, is implicated in disease pathophysiology.
- Oxygen regulation is critical for stem cell development, especially in early embryogenesis.
- Hypoxia-inducible factors (HIFs) are key regulators of cellular responses to hypoxia.
Purpose of the Study:
- To investigate the hypoxic regulation of HIF activity in placental trophoblast cells.
- To examine the impact of hypoxia on reporter gene expression in these cells.
- To evaluate the utility of reporter assays under hypoxic conditions.
Main Methods:
- Utilized dual-reporter luciferase assays in placental trophoblast cells.
- Exposed cells to controlled hypoxic conditions.
- Measured constitutive luciferase reporter activity.
Main Results:
- Hypoxic conditions led to a significant increase in constitutive luciferase reporter activity.
- This induction effect was observed across different cell types and species.
- Identified a potential method for normalizing transfection efficiency in hypoxic assays.
Conclusions:
- Hypoxia influences reporter gene expression in placental cells.
- Traditional dual-reporter assays may require re-evaluation under hypoxic conditions.
- Findings suggest caution in interpreting transcriptional activity measurements during hypoxia studies.
Related Concept Videos
01:23Hypoxia
Hypoxia is a medical condition characterized by an inadequate oxygen supply to body tissues. It typically manifests as a bluish discoloration of the skin and mucosae, especially in fair-skinned individuals, when hemoglobin (Hb) saturation drops below 75%.
Types of Hypoxia
There are four primary types of hypoxia, each resulting from a different cause:
1. Anemic hypoxia: This type occurs due to insufficient oxygen delivery caused by a lack of red blood cells (RBCs) or RBCs with abnormal or...
Types of Hypoxia
There are four primary types of hypoxia, each resulting from a different cause:
1. Anemic hypoxia: This type occurs due to insufficient oxygen delivery caused by a lack of red blood cells (RBCs) or RBCs with abnormal or...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...