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Ribosome Profiling02:24

Ribosome Profiling

3.6K
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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Protein-protein Interfaces02:04

Protein-protein Interfaces

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Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...
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Proteins: From Genes to Degradation02:11

Proteins: From Genes to Degradation

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Within a biological system, the DNA encodes the RNA, and the nucleotide sequence in the RNA further defines the amino acid sequence in the protein. This is referred to as “The Central Dogma of Molecular Biology” - a term coined by Francis Crick.  Central dogma is a firm principle in biology that defines the flow of genetic information within any life form. The two fundamental steps in central dogma are - transcription and translation.
Transcription is the synthesis of RNA...
12.8K
Leaky Scanning02:28

Leaky Scanning

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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.2K
Improving Translational Accuracy02:07

Improving Translational Accuracy

11.9K
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
11.9K
Translational Regulation01:29

Translational Regulation

104
Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
104

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Related Experiment Video

Updated: Sep 15, 2025

Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation
12:26

Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation

Published on: February 12, 2022

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Decoding RNA-Protein Interactions: Methodological Advances and Emerging Challenges.

Wenkai Yi1,2, Jian Yan1,2,3

  • 1Department of Biomedical Sciences Tung Biomedical Sciences Centre City University of Hong Kong Kowloon Tong Hong Kong.

Advanced Genetics (Hoboken, N.J.)
|July 14, 2025
PubMed
Summary
This summary is machine-generated.

This review compares RNA- and protein-centric methods for studying RNA-protein interactions, crucial for gene regulation. It guides researchers in selecting optimal techniques for advancing RNA biology and precision medicine.

Keywords:
Protein‐centric methodsRNARNA‐binding proteinsRNA‐centric methodsRNA–protein interactions

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An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA
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An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA

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iCLIP - Transcriptome-wide Mapping of Protein-RNA Interactions with Individual Nucleotide Resolution
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iCLIP - Transcriptome-wide Mapping of Protein-RNA Interactions with Individual Nucleotide Resolution

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iCLIP - Transcriptome-wide Mapping of Protein-RNA Interactions with Individual Nucleotide Resolution
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iCLIP - Transcriptome-wide Mapping of Protein-RNA Interactions with Individual Nucleotide Resolution

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • RNA-protein interactions are vital for cellular functions like gene regulation and RNA metabolism.
  • Recent methodological advancements offer enhanced resolution and specificity in studying these interactions.

Purpose of the Study:

  • To systematically compare RNA- and protein-centric methodologies for studying RNA-protein interactions.
  • To provide guidelines for selecting appropriate methods based on experimental goals and constraints.
  • To highlight challenges and future directions in the field.

Main Methods:

  • Comparison of RNA-centric approaches (e.g., pulldowns, proximity labeling, CRISPR-assisted techniques).
  • Evaluation of protein-centric strategies (e.g., CLIP-seq, proximity-tagging).
  • Assessment of innovative techniques like LACE-seq, ARTR-seq, and HyPro-MS.

Main Results:

  • RNA-centric methods identify proteins interacting with specific RNAs, including low-abundance partners.
  • Protein-centric methods map RNA interactomes with nucleotide precision.
  • Newer methods reduce cell input and bypass genetic modifications.

Conclusions:

  • Method selection should align with specific biological questions, RNA characteristics, and technical feasibility.
  • Addressing challenges like low-affinity interactions and RNA structure is key.
  • Tailored methodological approaches will accelerate discoveries in RNA biology and medicine.