Seeing Is Believing: Visualizing Circular RNAs

Advancement in the RNA sequencing techniques has discovered hundreds of thousands of circular RNAs (circRNAs) in humans. However, the physiological function of most of the identified circRNAs remains unexplored. Recent studies have established that spliceosomal machinery and RNA-binding proteins mod...

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Main Authors: Pruthvi Raj Bejugam, Aniruddha Das, Amaresh Chandra Panda
Format: Article
Language:English
Published: MDPI AG 2020-11-01
Series:Non-Coding RNA
Subjects:
Online Access:https://www.mdpi.com/2311-553X/6/4/45
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author Pruthvi Raj Bejugam
Aniruddha Das
Amaresh Chandra Panda
author_facet Pruthvi Raj Bejugam
Aniruddha Das
Amaresh Chandra Panda
author_sort Pruthvi Raj Bejugam
collection DOAJ
description Advancement in the RNA sequencing techniques has discovered hundreds of thousands of circular RNAs (circRNAs) in humans. However, the physiological function of most of the identified circRNAs remains unexplored. Recent studies have established that spliceosomal machinery and RNA-binding proteins modulate circRNA biogenesis. Furthermore, circRNAs have been implicated in regulating crucial cellular processes by interacting with various proteins and microRNAs. However, there are several challenges in understanding the mechanism of circRNA biogenesis, transport, and their interaction with cellular factors to regulate cellular events because of their low abundance and sequence similarity with linear RNA. Addressing these challenges requires systematic studies that directly visualize the circRNAs in cells at single-molecule resolution along with the molecular regulators. In this review, we present the design, benefits, and weaknesses of RNA imaging techniques such as single-molecule RNA fluorescence in situ hybridization and BaseScope in fixed cells and fluorescent RNA aptamers in live-cell imaging of circRNAs. Furthermore, we propose the potential use of molecular beacons, multiply labeled tetravalent RNA imaging probes, and Cas-derived systems to visualize circRNAs.
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spelling doaj.art-af3bbc03806d49a29f35b791bdf7eb8c2023-11-20T20:32:39ZengMDPI AGNon-Coding RNA2311-553X2020-11-01644510.3390/ncrna6040045Seeing Is Believing: Visualizing Circular RNAsPruthvi Raj Bejugam0Aniruddha Das1Amaresh Chandra Panda2Institute of Life Sciences, Nalco Square, Bhubaneswar, Odisha 751023, IndiaInstitute of Life Sciences, Nalco Square, Bhubaneswar, Odisha 751023, IndiaInstitute of Life Sciences, Nalco Square, Bhubaneswar, Odisha 751023, IndiaAdvancement in the RNA sequencing techniques has discovered hundreds of thousands of circular RNAs (circRNAs) in humans. However, the physiological function of most of the identified circRNAs remains unexplored. Recent studies have established that spliceosomal machinery and RNA-binding proteins modulate circRNA biogenesis. Furthermore, circRNAs have been implicated in regulating crucial cellular processes by interacting with various proteins and microRNAs. However, there are several challenges in understanding the mechanism of circRNA biogenesis, transport, and their interaction with cellular factors to regulate cellular events because of their low abundance and sequence similarity with linear RNA. Addressing these challenges requires systematic studies that directly visualize the circRNAs in cells at single-molecule resolution along with the molecular regulators. In this review, we present the design, benefits, and weaknesses of RNA imaging techniques such as single-molecule RNA fluorescence in situ hybridization and BaseScope in fixed cells and fluorescent RNA aptamers in live-cell imaging of circRNAs. Furthermore, we propose the potential use of molecular beacons, multiply labeled tetravalent RNA imaging probes, and Cas-derived systems to visualize circRNAs.https://www.mdpi.com/2311-553X/6/4/45circRNAFISHBaseScopeaptamerlocalizationRNA-binding protein
spellingShingle Pruthvi Raj Bejugam
Aniruddha Das
Amaresh Chandra Panda
Seeing Is Believing: Visualizing Circular RNAs
Non-Coding RNA
circRNA
FISH
BaseScope
aptamer
localization
RNA-binding protein
title Seeing Is Believing: Visualizing Circular RNAs
title_full Seeing Is Believing: Visualizing Circular RNAs
title_fullStr Seeing Is Believing: Visualizing Circular RNAs
title_full_unstemmed Seeing Is Believing: Visualizing Circular RNAs
title_short Seeing Is Believing: Visualizing Circular RNAs
title_sort seeing is believing visualizing circular rnas
topic circRNA
FISH
BaseScope
aptamer
localization
RNA-binding protein
url https://www.mdpi.com/2311-553X/6/4/45
work_keys_str_mv AT pruthvirajbejugam seeingisbelievingvisualizingcircularrnas
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