Optimized miR-124 reporters uncover differences in miR-124 expression among neuronal populations in vitro

Front Neurosci. 2023 Oct 18:17:1257599. doi: 10.3389/fnins.2023.1257599. eCollection 2023.

Abstract

Introduction: Although intensively studied in the last decades, how microRNAs (miRNAs) are expressed across different cell types in the brain remains largely unknown.

Materials: To address this issue, we sought to develop optimized fluorescence reporters that could be expressed in precise cellular subsets and used to accurately quantify miR contents in vivo.

Results: Focusing on miR-124, we tested different reporter designs whose efficiency was confirmed in different in vitro settings including cell lines and primary neuronal cultures from different brain structures. Unlike previous reporters, we provide experimental evidence that our optimized designs can faithfully translate miR levels in vitro.

Discussion: Tools developed here would enable assessing miRNA expression at the single cell resolution and are expected to significantly contribute to future miRNA research in vivo.

Keywords: AAV; miR-124 reporters; miRNA; mice; neuron.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the French National Agency (ANR) to EG (ANR-22-CE17-0034), Fondation de France (00100077), and Fondation France Alzheimer (2021-#6239).