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 the lack of technologies for probing them in live mouse brain. Here, we describe a novel method to visualize RNA granules in native CNS tissues in vivo. 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, a significant population of RNPs could be detected in specific cells of the CNS, enabling new findings, e.g., remarkable heterogeneity of Actb mRNA dynamics across neurons and glial cells. The highly sensitive in vivo RNA imaging could be useful for illuminating the regulation of the dynamics of RNA granules in naive and diseased animals.