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Related Concept Videos

FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

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Related Experiment Video

Updated: Jun 14, 2026

Bimolecular Fluorescence Complementation
08:54

Bimolecular Fluorescence Complementation

Published on: April 15, 2011

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Imaging Retroviral RNA Genome Heterodimers Using Bimolecular Fluorescence Complementation (BiFC).

Eunice C Chen1, Rebecca K Maldonado2,3, Leslie J Parent2,3

  • 1Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, Penn State College of Medicine, Hershey, PA 17033, USA.

Viruses
|August 28, 2025
PubMed
Summary

Researchers developed a novel imaging technique to detect retroviral genome heterodimers in living cells. This method uses modified bimolecular fluorescence complementation (BiFC) to visualize and track distinct viral RNA genomes interacting within virions.

Keywords:
RNA genome dimerRNA-RNA intermolecular interactionsRSVbimolecular fluorescence complementationconfocal microscopyfluorescence imagingretroviruses

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Retroviruses package two copies of their RNA genomes as dimers into virions.
  • Genome dimerization is crucial for retroviral infectivity and genetic diversity.
  • Retroviruses can package homodimers (identical RNAs) or heterodimers (distinct RNAs).

Purpose of the Study:

  • To develop a novel method for specifically labeling and detecting retroviral genome heterodimers in living cells.
  • To visualize and track the formation and dynamics of heterodimeric viral RNA genomes.
  • To investigate the interaction of heterodimeric genomes with viral proteins.

Main Methods:

  • Utilized a modified bimolecular fluorescence complementation (BiFC) technique.
  • Engineered viral genomes with distinct RNA stem-loop cassettes that bind specific RNA-binding proteins.
  • Conjugated split fluorophore components to RNA-binding proteins, allowing fluorescence reconstitution upon heterodimer proximity.

Main Results:

  • Successfully labeled and detected retroviral genome heterodimers in living cells using BiFC.
  • Visualized and tracked BiFC-labeled RNA dimers in real-time.
  • Demonstrated that these labeled heterodimers interact with retroviral Gag proteins.

Conclusions:

  • The developed BiFC-based imaging approach enables specific detection of retroviral genome heterodimers.
  • This method offers low background fluorescence and allows for dynamic studies of RNA-RNA interactions in living cells.
  • The technique is adaptable for studying dimeric or double-stranded RNAs in various biological systems, including other viruses and organisms.