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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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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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Transient Gene Expression in Tobacco using Gibson Assembly and the Gene Gun
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The Physcomitrella patens System for Transient Gene Expression Assays.

Johanne Thévenin1,2, Wenjia Xu1,2, Louise Vaisman1,2

  • 1INRA, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France.

Methods in Molecular Biology (Clifton, N.J.)
|August 26, 2016
PubMed
Summary

Transient expression assays using Physcomitrella patens protoplasts offer a fast and reliable method to study gene regulation. This technique combines green fluorescent protein (GFP) with flow cytometry for accurate analysis of transcriptional activity.

Keywords:
Physcomitrella patensProtoplastsSubcellular localizationTranscription factorTransient expression

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Area of Science:

  • Molecular Biology
  • Plant Science
  • Genetics

Background:

  • Transient expression assays are crucial for investigating gene expression regulation in vivo.
  • Understanding transcription factor (TF) binding to cis-regulatory elements is key to deciphering gene control.
  • Existing methods may lack speed, efficiency, or accuracy in measuring transcriptional properties.

Purpose of the Study:

  • To develop a rapid, efficient, and dependable method for studying gene expression.
  • To enable both qualitative and quantitative analysis of transcriptional regulation.
  • To utilize Physcomitrella patens protoplasts for in vivo gene expression studies.

Main Methods:

  • Employing Physcomitrella patens protoplasts for transient expression analysis.
  • Utilizing green fluorescent protein (GFP) as a reporter for promoter activity.
  • Implementing flow cytometry for precise, single-cell fluorescence measurement.

Main Results:

  • Demonstrated a fast and reliable method for assessing gene expression.
  • Enabled qualitative and quantitative analysis of promoter activity.
  • Achieved accurate measurement of fluorescence in individual cells via flow cytometry.

Conclusions:

  • The described method provides a powerful tool for studying transcriptional regulation.
  • Physcomitrella patens protoplast-based assays offer advantages in speed and accuracy.
  • This technique facilitates in-depth analysis of transcription factor function and cis-regulatory elements.