RNA Imaging pp 193-208 | Cite as

Positive Bioluminescence Imaging of MicroRNA Expression in Small Animal Models Using an Engineered Genetic-Switch Expression System, RILES

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1372)

Abstract

MicroRNAs (miRNAs) are a class of small, noncoding RNAs which regulate gene expression by directing their target mRNA for degradation or translational repression. Since their discovery in the early 1990s, miRNAs have emerged as key components in the posttranscriptional regulation of gene networks, shaping many biological processes from development, morphogenesis, differentiation, proliferation and apoptosis. Although understanding of the molecular basis of miRNA biology is improving, methods to monitor the dynamic and the spatiotemporal aspects of miRNA expression under physiopathological conditions are required. However, monitoring of miRNAs is difficult due to their small size, low abundance, high degree of sequence similarity, and their dynamic expression pattern which is subjected to tight transcriptional and post-transcriptional controls. Recently, we developed a miRNA monitoring system called RILES, standing for RNAi-inducible expression system, which relies on an engineered regulatable expression system, to switch on the expression of the luciferase gene when the targeted miRNA is expressed in cells. We demonstrated that RILES is a specific, sensitive, and robust method to determine the fine-tuning of miRNA expression during the development of an experimental pathological process in mice. Because RILES offers the possibility for longitudinal studies on individual subjects, sharper insights into miRNA regulation can be generated, with applications in physiology, pathophysiology and development of RNAi-based therapies. This chapter describes methods and protocols to monitor the expression of myomiR-206, -1, and -133 in the tibialis anterior muscle of mice. These protocols can be used and adapted to monitor the expression of other miRNAs in other biological processes.

Key words

microRNA Molecular imaging Bioluminescence study Spatiotemporal regulation RILES (RNAi-inducible luciferase expression system) 
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Notes

Acknowledgments

This work was supported by La ligue Contre le Cancer du Loiret, Région Centre. We would like thank the staff of the CIPA animal facility (TAAM/CIPA, CNRS UPS 44, Orléans) for their technical assistance and access to the Lumina Bioluminescence scanner. In addition, we would like to thank Dr. Lily Mijouin for helping in the preparation of the manuscript for this chapter.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Centre de Biophysique Moléculaire, CNRS UPR4301Université d’Orléans FranceOrléans Cedex 2France

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