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Related Concept Videos

RNA Interference01:23

RNA Interference

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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...
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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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Alternative RNA Splicing02:18

Alternative RNA Splicing

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
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RNA Splicing01:32

RNA Splicing

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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

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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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Types of RNA01:23

Types of RNA

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Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
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Single-cell RNA Sequencing and Analysis of Human Pancreatic Islets
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Introduction to Single-Cell RNA Sequencing.

Thale Kristin Olsen1, Ninib Baryawno1

  • 1Childhood Cancer Research Unit, Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.

Current Protocols in Molecular Biology
|June 1, 2018
PubMed
Summary

Single-cell RNA sequencing (scRNA-seq) offers unprecedented resolution for studying gene expression by analyzing individual cells, overcoming limitations of bulk RNA sequencing. This guide covers common scRNA-seq protocols, data analysis, and project design considerations.

Keywords:
RNA sequencinggene expression profilingsingle-cell analysis

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • High-throughput sequencing, particularly RNA sequencing (RNA-seq), has transformed biological research.
  • Bulk RNA-seq provides averaged gene expression data, potentially masking cell-to-cell variability.
  • Single-cell RNA sequencing (scRNA-seq) addresses this limitation by analyzing transcriptomes at the individual cell level.

Purpose of the Study:

  • To introduce common single-cell RNA sequencing (scRNA-seq) protocols.
  • To outline the fundamental principles of scRNA-seq data analysis.
  • To guide researchers in planning and designing effective scRNA-seq projects.

Main Methods:

  • Isolation of single cells for transcriptomic analysis.
  • Generation of sequencing libraries with transcripts mapped to individual cells.
  • Application of various scRNA-seq protocols and subsequent data processing.

Main Results:

  • scRNA-seq enables the assessment of biological properties at single-cell resolution.
  • Detailed gene expression patterns can be resolved within cell populations.
  • This technology facilitates a deeper understanding of biological systems.

Conclusions:

  • scRNA-seq provides a powerful approach to overcome the limitations of bulk RNA sequencing.
  • Understanding protocols and data analysis is crucial for successful scRNA-seq studies.
  • This method offers unprecedented resolution for biological discovery.