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Harnessing PUF-Based Reporters for Noninvasive Imaging of the MicroRNA Dynamics in Differentiation
- Source :
- Analytical Chemistry; March 2023, Vol. 95 Issue: 10 p4786-4794, 9p
- Publication Year :
- 2023
-
Abstract
- Precise characterization of miRNA expression patterns is critical to exploit the complexity of miRNA regulation in biology. Herein, we developed a Pumilio/FBF (PUF) protein-based engineering luciferase reporter system, PUF/miR, to quantitatively and non-invasively sense miRNA activity in living cells and animal models. We verified the feasibility of this reporter by monitoring the expression of several types of miRNAs (miRNA-9, 124a, 1, and 133a) in neural and muscle differentiated cells as well as subcutaneous or tibial anterior muscles in mice. The quantitative RT-PCR also validated the reliability and quantitative consistency of bioluminescence imaging in detecting miRNA expression. We further effectively employed this reporter system to visualize the expression of miRNA-1 and miRNA-133a in mouse models of skeletal muscle injury. As a non-invasive and convenient innovative approach, our results have realized the positive bioluminescence imaging of endogenous miRNAs in vitro and in vivo using the PUF/miR system. We believe that this approach would provide a potential means for noninvasive monitoring of disease-related miRNAs and could facilitate a deeper understanding of miRNA biology.
Details
- Language :
- English
- ISSN :
- 00032700 and 15206882
- Volume :
- 95
- Issue :
- 10
- Database :
- Supplemental Index
- Journal :
- Analytical Chemistry
- Publication Type :
- Periodical
- Accession number :
- ejs62379818
- Full Text :
- https://doi.org/10.1021/acs.analchem.3c00101