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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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Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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Related Experiment Video

Updated: Jun 16, 2025

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Analysis of Oligodendrocyte Lineage Cell Progression with Cre-Mediated RiboTag Reporter Lines.

George S Melchor1,2, Maya S Shah1, Zeeba Manavi1

  • 1Department of Biology, Georgetown University, Washington, DC, USA.

ASN Neuro
|June 14, 2025
PubMed
Summary

The RiboTag system effectively labels oligodendrocyte lineage cells in mice. Careful consideration of experimental models and timelines is crucial for accurate cell tracking and interpreting gene expression during development and injury.

Keywords:
Lineage tracingRiboTagoligodendrocyte precursor cellsoligodendrocytesremyelination

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Genetics

Background:

  • Cre-reporter strategies are vital for studying gene promoter activity and cell fate mapping in transgenic mice.
  • The RiboTag system utilizes a Cre/loxP-inducible hemagglutinin (HA)-tagged ribosomal protein (RPL22) for ribosome profiling.
  • Understanding RiboTag expression in oligodendrocyte lineage cells (OLCs) is essential for developmental and injury studies.

Purpose of the Study:

  • To profile RiboTag reporter expression in oligodendrocyte lineage cells (OLCs) using specific transgenic mouse models.
  • To evaluate the utility of RiboTag for tracking oligodendrocyte precursor cells (OPCs) and oligodendrocytes during development and demyelinating injury.
  • To assess the RiboTag system's sensitivity, stability, and suitability for various experimental conditions.

Main Methods:

  • Generation and analysis of NG2Cre:Rpl22HA, PdgfraCreERT:Rpl22HA, and PlpCreERT:Rpl22HA transgenic mice.
  • Assessment of HA reporter expression in the central nervous system (CNS) under basal conditions and following tamoxifen administration.
  • Evaluation of RiboTag labeling in OLCs in response to focal demyelinating injury and during remyelination.

Main Results:

  • NG2Cre:Rpl22HA mice showed HA expression in OLCs and neuronal populations.
  • PdgfraCreERT and PlpCreERT mice exhibited widespread HA labeling in OPCs and oligodendrocytes, respectively, upon tamoxifen induction.
  • RiboTag successfully labeled OPCs and oligodendrocytes during remyelination, but PlpCreERT labeling complicated the distinction between new and pre-existing oligodendrocytes.
  • A notable observation in PlpCreERT mice was the induction of HA expression in oligodendrocytes but not OPCs in demyelinated lesions, suggesting post-transcriptional regulation.

Conclusions:

  • The RiboTag reporter system demonstrates high sensitivity and stability for labeling OLCs.
  • The choice of Cre driver and experimental timeline significantly impacts the interpretation of RiboTag data.
  • Careful consideration of the RiboTag system's application is necessary, taking into account the specific experimental model, timing, and cell tracking requirements.