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RNA-seq03:21

RNA-seq

10.0K
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. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

16.8K
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
16.8K
Nucleic Acid Structure01:25

Nucleic Acid Structure

6.2K
The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms  a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA...
6.2K
Ribosome Profiling02:24

Ribosome Profiling

3.5K
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...
3.5K
RNA Interference01:23

RNA Interference

26.1K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
26.1K
RNA Stability01:53

RNA Stability

33.6K
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...
33.6K

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Related Experiment Video

Updated: Jul 9, 2025

Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs
08:49

Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs

Published on: September 16, 2019

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Small RNA structural biochemistry in a post-sequencing era.

Juan Pablo Tosar1,2, Mauricio Castellano3,4, Bruno Costa3,5

  • 1Functional Genomics Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. jptosar@pasteur.edu.uy.

Nature Protocols
|December 6, 2023
PubMed
Summary

Small RNA sequencing offers a limited view of the transcriptome, often introducing biases and losing crucial 3D structural information. Integrating other techniques is vital for accurate RNA fragmentomics interpretation.

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Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells
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Author Spotlight: AQRNA-seq Role in Mapping Small RNAs and Unraveling Protein Translation Mechanisms
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Author Spotlight: AQRNA-seq Role in Mapping Small RNAs and Unraveling Protein Translation Mechanisms

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Author Spotlight: AQRNA-seq Role in Mapping Small RNAs and Unraveling Protein Translation Mechanisms
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Author Spotlight: AQRNA-seq Role in Mapping Small RNAs and Unraveling Protein Translation Mechanisms

Published on: February 2, 2024

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • High-throughput sequencing significantly advanced small RNA research but has inherent limitations.
  • Current 'sequence, map and annotate' methods can yield biologically implausible results due to biases and database issues.
  • Loss of 3D structural information hinders understanding of RNA stability, turnover, and function.

Purpose of the Study:

  • To discuss sources of bias in small RNA sequencing.
  • To present strategies for reducing bias in sequencing data.
  • To provide guidance on interpreting small RNA sequencing results, focusing on RNA fragmentomics.

Main Methods:

  • Review of common biases in high-throughput sequencing of small RNAs.
  • Discussion of bias-reducing strategies.
  • Emphasis on integrating structural knowledge and complementary techniques.

Main Results:

  • Sequencing provides a 1D view, potentially missing 3D structural aspects like RNA fragment dimers or nicked forms.
  • Database inaccuracies and inherent sequencing biases can lead to flawed interpretations.
  • Loss of structural information is a major limitation in understanding RNA dynamics.

Conclusions:

  • Small RNA sequencing requires careful interpretation due to inherent biases and loss of dimensional information.
  • Strategies to mitigate bias and integrate structural data are crucial for accurate RNA fragmentomics.
  • Combining sequencing with orthologous experimental techniques is strongly recommended for robust biological insights.