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Related Concept Videos

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

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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq
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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq

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Identifying Trypanosome Protein-RNA Interactions Using RIP-Seq.

Elisha Mugo1, Esteban D Erben2

  • 1Department of Biochemistry, Genetics and Microbiology, Institute for Sustainable Malaria Control, University of Pretoria, Hatfield, South Africa.

Methods in Molecular Biology (Clifton, N.J.)
|March 30, 2020
PubMed
Summary

This study details a new RNA immunoprecipitation sequencing (RIP-Seq) protocol to map RNA-binding protein interactions in Trypanosoma brucei. This method helps understand gene expression control in trypanosomatids.

Keywords:
Protein characterizationRNA Immunoprecipitation (RIP)RNA-binding proteinsmRNA target identification

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Area of Science:

  • Molecular Biology
  • Genetics
  • Parasitology

Background:

  • Trypanosomatids primarily use posttranscriptional mechanisms to regulate gene expression.
  • RNA-binding proteins (RBPs) are key regulators of RNA processing, localization, degradation, and translation in these organisms.

Purpose of the Study:

  • To identify mRNA targets of RBPs across the transcriptome.
  • To map the specific RNA-binding sites of RBPs.
  • To elucidate the mechanisms of gene expression control by RBPs in trypanosomatids.

Main Methods:

  • RNA immunoprecipitation followed by high-throughput sequencing (RIP-Seq) protocol.
  • Application of the protocol to the model organism Trypanosoma brucei.

Main Results:

  • The study presents an updated RIP-Seq protocol for analyzing RNA/protein interactions.
  • The technique allows for transcriptome-wide identification of mRNA targets.
  • Mapping of RNA-binding sites provides insights into RBP function.

Conclusions:

  • The developed RIP-Seq protocol is valuable for studying messenger ribonucleoprotein (mRNP) networks.
  • This method enhances our understanding of gene expression mechanisms in trypanosomes.