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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

4.1K
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...
4.1K

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Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling
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A simple, fast, and cost-efficient protocol for ultra-sensitive ribosome profiling.

Jiří Koubek1, Katharina Jetzinger1, Shiran Dror1

  • 1Center for Molecular Biology of Heidelberg University (ZMBH) and German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, Heidelberg, 69120, Germany.

Nucleic Acids Research
|September 18, 2025
PubMed
Summary
This summary is machine-generated.

We developed a cost-effective ribosome profiling method using bead-coupled reactions. This ultra-sensitive protocol enhances yield and throughput for messenger RNA (mRNA) translation studies, even with minimal RNA input.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • Ribosome profiling is crucial for studying messenger RNA (mRNA) translation at codon resolution.
  • Current methods are limited by labor intensity, low efficiency, and high costs.
  • There is a need for more accessible and efficient ribosome profiling techniques.

Purpose of the Study:

  • To present a novel, cost-effective, and ultra-sensitive library preparation method for ribosome profiling.
  • To improve the yield, throughput, and reproducibility of ribosome profiling workflows.
  • To expand the applicability of ribosome profiling to minimal input samples.

Main Methods:

  • Implementation of bead-coupled enzymatic reactions for library preparation.
  • Utilized product purifications to enhance yield and throughput.
  • Tested the protocol with as little as 12 fmol of RNA.

Main Results:

  • Achieved significantly increased yield and throughput with high reproducibility.
  • Demonstrated ultra-sensitivity, enabling ribosome profiling from minimal RNA input (12 fmol).
  • Validated the protocol's versatility across multiple species and for RNA-sequencing library preparation.

Conclusions:

  • The developed protocol offers a highly accessible and efficient alternative for ribosome profiling.
  • Facilitates ribosome profiling in challenging experimental contexts, including small cell populations and patient samples.
  • Advances the study of mRNA translation with codon-level resolution.