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Related Concept Videos

lncRNA - Long Non-coding RNAs02:39

lncRNA - Long Non-coding RNAs

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In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...
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siRNA - Small Interfering RNAs02:30

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PIWI-interacting RNAs, or piRNAs, are the most abundant short non-coding RNAs. More than 20,000 genes have been found in humans that code for piRNAs while only 2000 genes have been found for miRNAs. piRNAs can act at the transcriptional and post-transcriptional levels and have a vital role in silencing transposable elements present in germ cells. They are also involved in epigenetic silencing and activation. Previously, they were thought to function only in germ cells but new evidence suggests...
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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.
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One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
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Related Experiment Video

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Determination of In Vitro and Cellular Turn-on Kinetics for Fluorogenic RNA Aptamers
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Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells.

Alexis Autour1, Sunny C Y Jeng2, Adam D Cawte3,4

  • 1Université de Strasbourg, CNRS, Architecture et Réactivité de l'ARN, UPR 9002, 67000, Strasbourg, France.

Nature Communications
|February 15, 2018
PubMed
Summary

Researchers developed new fluorescent RNA Mango aptamers for imaging RNA in cells. These tools are bright, preserve RNA function, and enable visualization of small non-coding RNAs in live and fixed mammalian cells.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • Directly imaging biologically important RNAs in cells is challenging due to a lack of suitable fluorescent tools.
  • Existing protein imaging tools, like fluorescent proteins, are not directly applicable to RNA studies.
  • Ideal RNA labeling systems require high affinity, preservation of biological function, and favorable photophysical properties.

Purpose of the Study:

  • To develop novel fluorescent aptamers for high-resolution RNA imaging in cellular environments.
  • To create RNA labeling tools with photophysical properties comparable to existing fluorescent proteins.
  • To enable the study of RNA localization and dynamics in live and fixed mammalian cells.

Main Methods:

  • Microfluidics-based selection was employed to identify high-affinity RNA Mango fluorogenic aptamers.
  • The photophysical properties of the new aptamers were characterized, including brightness compared to enhanced GFP.
  • The aptamers were used to visualize the subcellular localization of specific small non-coding RNAs in mammalian cells.

Main Results:

  • Three new high-affinity RNA Mango fluorogenic aptamers were successfully selected.
  • Two of the developed aptamers demonstrated brightness equal to or exceeding enhanced GFP when bound to TO1-Biotin.
  • Accurate imaging of subcellular localization for 5S, U6, and box C/D scaRNA was achieved in both live and fixed mammalian cells.

Conclusions:

  • The newly developed RNA Mango aptamers offer a significant advancement for RNA visualization in biological systems.
  • These aptamers provide a powerful new tool for studying RNA function, dynamics, and localization in vitro and in mammalian cells.
  • The developed tools are compatible with established live and fixed cell protein labeling strategies, facilitating multi-modal imaging.