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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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Fluorescent mRNA labeling through cytoplasmic FISH.

Maxime Gasnier1, Cynthia Dennis, Catherine Vaurs-Barrière

  • 11] INSERM, U1103, Laboratoire Génétique, Reproduction et Développement (GreD), Clermont-Ferrand, France. [2] CNRS, UMR6293, Laboratoire GReD, Clermont-Ferrand, France. [3] Clermont Université, Laboratoire GReD, Clermont-Ferrand, France. [4].

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Summary
This summary is machine-generated.

Cytoplasmic fluorescence in situ hybridization (cFISH) allows detailed RNA visualization in cells and tissues. This method enables multiple RNA probe and antibody labeling for enhanced gene expression studies in mouse embryos.

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Genetics

Background:

  • RNA in situ hybridization (ISH) is crucial for studying gene expression in biological research.
  • Traditional colorimetric ISH methods, like NBT/BCIP, lack multi-probe capability and cellular resolution.

Purpose of the Study:

  • To introduce a novel cytoplasmic fluorescence in situ hybridization (cFISH) protocol.
  • To enable high-resolution RNA visualization from whole tissues down to single cells.

Main Methods:

  • Developed a cFISH protocol adapted for mouse embryos.
  • Enabled multi-labeling by combining RNA probes and antibodies (immuno-cFISH).

Main Results:

  • cFISH provides visualization of specific RNA strands at cellular resolution.
  • The protocol supports multiplexing of RNA probes and antibodies for comprehensive analysis.

Conclusions:

  • cFISH offers a powerful, high-resolution alternative to classical ISH.
  • The protocol can be completed within 2-3 days, facilitating rapid gene expression analysis.