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Related Concept Videos

Regulated mRNA Transport02:22

Regulated mRNA Transport

In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...
Nuclear Localization Signals and Import01:46

Nuclear Localization Signals and Import

Proteins targeted to the nucleus carry short stretches of amino acid sequences called the nuclear localization signal or NLS. Classical nuclear localization signals are of two types: monopartite and bipartite NLS. Monopartite classical NLS (cNLS) consists of a single cluster of 4-8 amino acids. Bipartite cNLS consists of two clusters of  2-3 amino acids and a 9-12 residue long proline-rich linker bridging the two clusters. Signal clusters are rich in positively charged amino acids such as...

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Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
11:24

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Published on: June 17, 2015

Neuropeptide RNA localization in tissue sections.

Marc Landry1, Shérine Abdel Salam, Marie Moftah

  • 1INSERM U862, University of Bordeaux, Bordeaux, France. marc.landry@u-bordeaux2.fr

Methods in Molecular Biology (Clifton, N.J.)
|September 17, 2011
PubMed
Summary
This summary is machine-generated.

In situ hybridization is a versatile technique for visualizing RNA in cells. Protocols vary based on RNA type (mRNA, miRNA) and detection method (microscopy, immunocytochemistry), requiring careful selection for specific research needs.

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Localized RNAi and Ectopic Gene Expression in the Medicinal Leech
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Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
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Published on: June 17, 2015

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • In situ hybridization (ISH) is a standard method for determining RNA localization within cells and tissues.
  • Numerous ISH protocols exist, necessitating clear definition of research objectives for optimal method selection.
  • ISH can detect various RNA types, including messenger RNA (mRNA) and small noncoding RNAs like microRNA (miRNA).

Purpose of the Study:

  • To present diverse in situ hybridization protocols for RNA detection.
  • To highlight the adaptability of ISH for different RNA targets and microscopy techniques.
  • To guide researchers in selecting appropriate ISH methods based on specific experimental requirements.

Main Methods:

  • Detailed protocols for in situ hybridization are presented.
  • Methods cover the detection of both mRNA and miRNA.
  • Protocols are adaptable for light and electron microscopy and can be combined with immunocytochemistry.

Main Results:

  • In situ hybridization protocols show significant diversity depending on the application.
  • Detection of mRNA and miRNA utilizes similar core protocols but requires specific probe types.
  • Oligo- and riboprobes, along with locked nucleic-acid probes, are crucial for specific RNA detection.

Conclusions:

  • The choice of in situ hybridization protocol is critical and depends on the specific RNA target and detection requirements.
  • While core ISH methods are similar for mRNA and miRNA, probe selection is key.
  • ISH is a powerful technique for studying RNA localization and coexpression with proteins.