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Related Concept Videos

Reporter Genes02:11

Reporter 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...

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Related Experiment Video

Updated: Jun 19, 2026

Quantitative Measurement of Relative Retinoic Acid Levels in E8.5 Embryos and Neurosphere Cultures Using the F9 RARE-Lacz Cell-based Reporter Assay
09:49

Quantitative Measurement of Relative Retinoic Acid Levels in E8.5 Embryos and Neurosphere Cultures Using the F9 RARE-Lacz Cell-based Reporter Assay

Published on: September 6, 2016

Multiplexed Retinoic Acid Signal Reporters for In Vivo and In Vitro Use.

Craig Smith1,2, Bianca Black3, Ric Broadhurst1

  • 1Reproductive Technologies, AgResearch Ruakura, Hamilton, New Zealand.

Genesis (New York, N.Y. : 2000)
|June 18, 2026
PubMed
Summary
This summary is machine-generated.

Researchers optimized a modular reporter system for detecting retinoic acid (RA) levels. This method uses dual reporters and insulators for accurate measurement of RA concentration and context-dependent responses in embryos.

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Genetics

Background:

  • Developing sensitive reporters for signaling molecules like retinoic acid (RA) is crucial for understanding developmental processes.
  • Existing methods for generating transgenic models can be complex, often requiring embryonic stem cells.

Purpose of the Study:

  • To optimize a modular, adaptable, and rapid method for creating multimerized signaling-molecule reporters.
  • To enable the detection of varying retinoic acid concentrations and study its effects in vivo.
  • To develop a system that avoids the use of embryonic stem cells for generating transgenic lines.

Main Methods:

  • Designed a modular plasmid system allowing easy exchange of reporters, promoters, and signal-responsive elements.
  • Utilized pronuclear injection for random integration into genomic DNA, bypassing embryonic stem cells.
  • Incorporated insulator elements to prevent integration-specific influences and cassette-to-cassette interference.
  • Tested varying numbers of retinoic acid receptor binding sites (2, 4, 12 copies) to assess sensitivity.
  • Employed dual reporters to control for confounding integration and copy-number dependent effects.

Main Results:

  • The modular reporter system demonstrated efficient detection of retinoic acid concentrations.
  • Insulator elements improved ligand concentration responsiveness in transient transfections and transgenic embryos.
  • Increasing binding sites enhanced in vitro RA sensitivity more than in vivo sensitivity.
  • Robust expression patterns were observed in specific embryonic tissues, including the primitive streak and somites.
  • Dual reporters effectively controlled for integration and copy-number effects, revealing context-dependent responses.

Conclusions:

  • The optimized modular reporter system provides a versatile tool for studying signaling pathways, specifically retinoic acid.
  • The use of dual reporters with insulators is effective for concurrently measuring concentration and context-dependent responses.
  • This approach facilitates the examination of interactions between multiple signaling pathways within a single embryo.