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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...
DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
Ribosome Profiling02:24

Ribosome Profiling

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 helps...
Diversity in Cell Signaling Responses01:22

Diversity in Cell Signaling Responses

The physiological function of a cell and cellular communication are outcomes of a range of extrinsic signals, intracellular signaling pathways, and cellular responses. No two cell types express the same repertoire of signaling components. Receptors are highly selective for their cognate ligands, but once activated, they can alter multiple cellular processes such as DNA transcription, protein synthesis, and metabolic activity. 
Graded and Abrupt Responses
Some signaling systems generate...

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Related Experiment Video

Updated: May 29, 2026

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
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Published on: July 3, 2013

Linking cellular signalling to gene expression using EXT-encoded reporter libraries.

Anna Botvinik1, Moritz J Rossner

  • 1Research Group 'Gene Expression', Max-Planck-Institute of Experimental Medicine, Goettingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|September 23, 2011
PubMed
Summary

This study introduces the expressed oligonucleotide tag (EXT) assay, a novel method for analyzing cellular signaling pathways upstream of gene expression. This technique allows for parallel analysis of complex signaling events in living cells.

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Area of Science:

  • Cellular biology
  • Molecular biology
  • Biochemistry

Background:

  • Intracellular signaling involves complex, interconnected biochemical steps initiated by cell surface receptors.
  • Protein-protein interactions, regulated by kinases and phosphorylation, are crucial for signaling cascades.
  • Studying transcriptional changes is feasible, but analyzing upstream regulatory mechanisms remains challenging.

Purpose of the Study:

  • To develop a novel experimental approach for parallel analysis of signaling events upstream of gene expression.
  • To overcome the challenges in measuring upstream regulatory mechanisms in cellular signaling.

Main Methods:

  • Developed the expressed oligonucleotide tag (EXT) assay.
  • Utilized reporter gene assays linked to unique EXTs as quantitative decoders.
  • Introduced EXT reporters into living cells for pooled analysis via microarray hybridization or sequencing.

Main Results:

  • The EXT assay enables parallel analysis of signaling events upstream of gene expression.
  • Reporter gene assays linked to EXTs serve as quantitative decoders for signaling events.
  • Pooled analysis of EXT reporters in living cells is achievable through established high-throughput methods.

Conclusions:

  • The EXT assay provides a powerful new tool for dissecting complex cellular signaling pathways.
  • This method facilitates the study of upstream regulatory mechanisms, complementing existing transcriptional analysis techniques.
  • The EXT assay allows for quantitative decoding of signaling events, advancing the understanding of cellular responses.