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Updated: Apr 23, 2026

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications

Published on: July 10, 2020

In vivo RNA labeling using MS2.

Eduardo Peña¹, Manfred Heinlein, Adrian Sambade

¹Institut de Biologie Moléculaire des Plantes (IBMP), Centre National de la Recherche Scientifique (CNRS), 12 rue du Général Zimmer, 67084, Strasbourg, France.

Methods in Molecular Biology (Clifton, N.J.)

|October 8, 2014

Summary

This summary is machine-generated.

Plant cells transport RNA between cells via plasmodesmata (PD), crucial for development and disease. This study details a method to visualize RNA trafficking in vivo using the MS2 system in Nicotiana benthamiana.

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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Published on: December 21, 2017

Area of Science:

Molecular Biology
Plant Biology
Cell Biology

Background:

Cytoplasmic RNA trafficking and asymmetric distribution are vital for plant development and signaling.
Plasmodesmata (PD) mediate intercellular RNA transport, impacting cell-to-cell communication and viral spread.
Advances in live-cell imaging enable molecular-level studies of dynamic biological processes.

Purpose of the Study:

To describe a protocol for in vivo visualization of transiently expressed RNA.
To investigate RNA localization and trafficking to and through plasmodesmata.
To leverage the MS2 tagging system for studying RNA dynamics in plants.

Main Methods:

Utilizing the MS2 method for RNA tagging with bacteriophage MS2 coat protein (CP) and fluorescent proteins.
Employing time-lapse microscopy for real-time monitoring of RNA distribution in living cells.
Visualizing SL-tagged RNA in vivo within Nicotiana benthamiana tissues.

Main Results:

Successful in vivo visualization of transiently expressed SL-tagged RNA in plants.
Demonstration of RNA localization and trafficking dynamics related to plasmodesmata.
Establishment of a robust method for studying RNA transport in a model plant system.

Conclusions:

The MS2 system provides a powerful tool for visualizing and studying RNA trafficking in plants.
Understanding RNA transport through plasmodesmata is key to deciphering plant development and disease mechanisms.
This protocol facilitates further research into the complex regulation of intercellular RNA movement.