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Related Concept Videos

What is Gene Expression?01:42

What is Gene Expression?

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Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
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What is Gene Expression?01:36

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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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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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Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
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Chromatin Position Affects Gene Expression

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Chromatin is the massive complex of DNA and proteins packaged inside the nucleus. The complexity of chromatin folding and how it is packaged inside the nucleus greatly influences  access to genetic information. Generally, the nucleus' periphery is considered transcriptionally repressive, while the cell's interior is considered a transcriptionally active area. 
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In vivo Dual Substrate Bioluminescent Imaging
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A Firefly Luciferase Dual Color Bioluminescence Reporter Assay Using Two Substrates To Simultaneously Monitor Two

Bruce R Branchini1, Tara L Southworth2, Danielle M Fontaine2

  • 1Department of Chemistry, Connecticut College, New London, Connecticut, 06320, USA. brbra@conncoll.edu.

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|April 18, 2018
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Summary

This study introduces a novel bioluminescence reporter assay for simultaneously monitoring two genes using distinct green and red light signals. The method offers improved background, cell normalization, and detectability for gene expression studies.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Bioluminescence is a sensitive method for monitoring eukaryotic gene expression.
  • Current methods often rely on single reporter genes and exogenous substrate addition.
  • Simultaneous monitoring of multiple genes can be challenging.

Purpose of the Study:

  • To develop a novel bioluminescence reporter method for simultaneous monitoring of two genes.
  • To engineer firefly luciferase variants with distinct substrate specificities for differential light emission (green and red).
  • To compare the performance of the new assay with existing methods.

Main Methods:

  • Engineered firefly luciferase variants emitting green and red light were developed.
  • The method utilizes selective luciferin substrates for each enzyme variant.
  • Experiments involved promoter transactivation assays and comparative analyses with the Dual-Luciferase Reporter protocol.

Main Results:

  • The novel method successfully monitored two genes simultaneously with distinguishable green and red light signals.
  • Results from promoter transactivation experiments were comparable to the Dual-Luciferase Reporter protocol.
  • The new assay demonstrated improvements in background noise, cell normalization, and signal detectability.

Conclusions:

  • This novel bioluminescence assay enables sensitive, simultaneous monitoring of two genes.
  • The method offers advantages over existing reporter assays, including simplified reagent handling.
  • It is a valuable tool for genetic reporter assays requiring dual-gene expression analysis.