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Related Concept Videos

Biological Clocks and Seasonal Responses02:45

Biological Clocks and Seasonal Responses

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The circadian—or biological—clock is an intrinsic, timekeeping, molecular mechanism that allows plants to coordinate physiological activities over 24-hour cycles called circadian rhythms. Photoperiodism is a collective term for the biological responses of plants to variations in the relative lengths of dark and light periods. The period of light-exposure is called the photoperiod.
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Reporting and recording are crucial in data documentation. The timely, thorough, and accurate documentation of facts is essential when recording patient data. Failure to record findings during an assessment or interpretation of a problem will result in loss of information and make the patient document unreliable. The reader is left with general impressions if the information is not specific. A recording is documenting data of the individual's health information in a traceable, secure, and...
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Types of Reports I: Hands-off Report01:25

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An Incident or Occurrence Report in a healthcare setting is a crucial document used to record any unexpected occurrence that may or may not have affected a patient, employee, or visitor. Such reports are critical to improving patient safety and include all details leading up to and including the event.
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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.
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Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters
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Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters

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Developing Mammalian Cellular Clock Models Using Firefly Luciferase Reporter.

Chidambaram Ramanathan1, Andrew C Liu2,3

  • 1Department of Biological Sciences, The University of Memphis, Memphis, TN, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 20, 2018
PubMed
Summary
This summary is machine-generated.

Mammalian circadian clocks regulate physiological processes via rhythmic gene expression. Researchers developed a stable mouse hepatocyte reporter cell line using luciferase bioluminescence for precise circadian parameter measurement.

Keywords:
BioluminescenceCircadian clockFirefly luciferaseHepatocytesMammalianPer2

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Area of Science:

  • Chronobiology
  • Molecular Biology
  • Cell Biology

Background:

  • Endogenous circadian clocks regulate mammalian metabolic, physiological, and behavioral processes.
  • Cellular and molecular mechanisms underlie rhythmic gene expression across different tissue types.
  • Individual cells function as rhythm generators, making cell-based clock models experimentally tractable.

Purpose of the Study:

  • To describe a procedure for establishing a stable mouse hepatocyte reporter cell line.
  • To demonstrate a method for creating cellular clock models using noninvasive bioluminescence reporters.
  • To enable precise determination of circadian parameters through high-temporal-resolution measurements.

Main Methods:

  • Fusion of rhythmically expressed gene promoters with the firefly luciferase (Luc) gene to create reporter constructs.
  • Development of stable reporter cell lines, specifically a mouse hepatocyte line.
  • Continuous, sensitive, and automated measurement of bioluminescence over several days.

Main Results:

  • Establishment of a stable mouse hepatocyte reporter cell line.
  • Generation of high-temporal-resolution bioluminescence data.
  • Precise determination of key circadian parameters: period length, amplitude, damping rate, and phase.

Conclusions:

  • Cellular clock models utilizing luciferase reporters provide a robust method for studying circadian rhythms.
  • The described procedure is applicable to various cell types beyond hepatocytes.
  • Miniaturization of these assays enhances throughput for large-scale screening of circadian clock function.