Amplified MS2 labeling reveals heterogeneous dynamics of RNA granules in the live murine brain ex vivo
- 1School of Optometry, Indiana University, Bloomington, Indianapolis, United States of America
Abstract
Dynamics of messenger ribonucleic acids (mRNAs) and the complexes with associated proteins (RNPs), also known as RNA granules, provide post-transcriptional control of gene expression. This regulation is crucial for cell-type diversity and activity-dependent plasticity of the mammalian central nervous system (CNS). However, elucidating the physiological significance of RNA granules has been hampered by technical difficulties to probe them in live brain. Here, we describe a novel method to visualize RNA granules in native CNS tissues of mouse. To amplify the fluorescence signal from single mRNAs incorporating MS2 stem loops, MS2 capsid protein (MCP) was conjugated with 4 tandem superfolder green fluorescent proteins (sfGFPs). Using MCP-4 × sfGFP, significantly more RNPs could be detected in the fresh acute brain slices than with MCP-GFP. Amplified MS2 signal could visualize Actb mRNAs in specific cells, either driven by promoter or via Cre-dependent expression, revealing remarkably distinct localization and movements of RNPs across different CNS cell types, including neurons and glial cells. The highly sensitive live-tissue RNA imaging could be useful for investigating the regulation of the dynamics of RNA granules in the native niche, ultimately in naive and diseased animal models.
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BibTeX
@article{lim2026amplified,
title = {{Amplified MS2 labeling reveals heterogeneous dynamics of RNA granules in the live murine brain ex vivo}},
author = {Hyungsik Lim and Grace P. Cooper},
journal = {PLOS One},
year = {2026},
month = nov,
doi = {10.1371/journal.pone.0361966},
url = {https://doi.org/10.1371/journal.pone.0361966},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Amplified MS2 labeling reveals heterogeneous dynamics of RNA granules in the live murine brain ex vivo
AU - Hyungsik Lim
AU - Grace P. Cooper
PY - 2026
DA - 2026/11/05
JO - PLOS One
DO - 10.1371/journal.pone.0361966
UR - https://doi.org/10.1371/journal.pone.0361966
AB - Dynamics of messenger ribonucleic acids (mRNAs) and the complexes with associated proteins (RNPs), also known as RNA granules, provide post-transcriptional control of gene expression. This regulation is crucial for cell-type diversity and activity-dependent plasticity of the mammalian central nervous system (CNS). However, elucidating the physiological significance of RNA granules has been hampered by technical difficulties to probe them in live brain. Here, we describe a novel method to visualize RNA granules in native CNS tissues of mouse. To amplify the fluorescence signal from single mRNAs incorporating MS2 stem loops, MS2 capsid protein (MCP) was conjugated with 4 tandem superfolder green fluorescent proteins (sfGFPs). Using MCP-4 × sfGFP, significantly more RNPs could be detected in the fresh acute brain slices than with MCP-GFP. Amplified MS2 signal could visualize Actb mRNAs in specific cells, either driven by promoter or via Cre-dependent expression, revealing remarkably distinct localization and movements of RNPs across different CNS cell types, including neurons and glial cells. The highly sensitive live-tissue RNA imaging could be useful for investigating the regulation of the dynamics of RNA granules in the native niche, ultimately in naive and diseased animal models.
SN - 1932-6203
ER -