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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

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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...
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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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What is Gene Expression?01:42

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Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
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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.
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Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
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What have single-molecule studies taught us about gene expression?

Huimin Chen1, Daniel R Larson1

  • 1Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

Genes & Development
|September 8, 2016
PubMed
Summary

Single-molecule techniques provide sensitive, real-time insights into gene expression. These methods reveal complex molecular mechanisms in vitro and in vivo, advancing our understanding of RNA production.

Keywords:
fluorescencesingle moleculesplicingtranscription

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Gene expression involves sequential steps: transcription factor docking, polymerase initiation, elongation, splicing, and termination.
  • Traditional knowledge relies on ensemble in vitro biochemical measurements.
  • Single-molecule approaches offer enhanced sensitivity and the ability to observe heterogeneous behaviors.

Purpose of the Study:

  • To review the unique insights gained from single-molecule techniques in studying gene expression mechanisms.
  • To highlight the advantages of single-molecule methods over traditional ensemble measurements.
  • To discuss the application of these techniques in both in vitro and in vivo systems.

Main Methods:

  • Single-molecule imaging and manipulation techniques.
  • Real-time observation of macromolecular dynamics.
  • In vitro biochemical assays.
  • In vivo studies in living cells.

Main Results:

  • Single-molecule methods reveal heterogeneity in gene expression processes.
  • These techniques allow direct observation of macromolecular interactions and dynamics.
  • Insights into transcription, RNA processing, and termination have been significantly advanced.
  • In vivo single-molecule studies address biological questions in their native cellular context.

Conclusions:

  • Single-molecule techniques are powerful tools for dissecting the complex mechanisms of gene expression.
  • They provide unprecedented resolution in space and time, complementing traditional biochemical methods.
  • The application of these methods in vivo opens new avenues for understanding gene regulation in living systems.