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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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RNA Structure01:23

RNA Structure

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Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
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Ribosomal RNA Synthesis02:53

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Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
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RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
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Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

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RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
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RNA Stability01:53

RNA Stability

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Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
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Related Experiment Video

Updated: May 30, 2025

Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells
10:34

Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells

Published on: December 9, 2022

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Consistent features observed in structural probing data of eukaryotic RNAs.

Kazuteru Yamamura1, Kiyoshi Asai1, Junichi Iwakiri1

  • 1Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 277-8561, Japan.

NAR Genomics and Bioinformatics
|January 31, 2025
PubMed
Summary
This summary is machine-generated.

Analyzing RNA chemical probing data in eukaryotes reveals key insights. High reactivity indicates solvent exposure and known secondary structures, but low reactivity does not reliably show solvent exposure.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genomics

Background:

  • Understanding RNA structure is vital for its regulatory functions and therapeutic applications, such as mRNA vaccines.
  • Messenger RNA (mRNA) structure significantly influences RNA stability and translation efficiency.
  • Chemical probing is a key experimental method for studying RNA structure within living cells.

Purpose of the Study:

  • To analyze transcriptome-scale RNA chemical probing data in eukaryotes.
  • To identify common features and potential biases in RNA chemical probing data.
  • To improve the interpretation of RNA chemical probing experiments.

Main Methods:

  • Comprehensive analysis of existing transcriptome-scale RNA chemical probing datasets from eukaryotic organisms.
  • Statistical evaluation of reactivity patterns in relation to known RNA secondary structures and solvent accessibility.
  • Identification of common trends and characteristics across multiple experimental datasets.

Main Results:

  • A small fraction of bases were modified in most probing experiments.
  • The top 10% most reactive bases accurately reflected known RNA secondary structures.
  • Highly reactive bases generally corresponded to solvent-exposed regions, while low reactivity did not reliably indicate solvent exposure.

Conclusions:

  • RNA chemical probing is a valuable tool for inferring RNA structure and dynamics in cells.
  • Awareness of data biases, particularly regarding low reactivity, is crucial for accurate interpretation.
  • These findings provide essential guidelines for analyzing RNA chemical probing data to better understand RNA structure-function relationships.