Protocol abstract
Nature Protocols 4, - 1663 - 1669 (2009)
Published online: 29 October 2009 | doi:10.1038/nprot.2009.119
Subject Categories: Genetic modification | Imaging
A reporter gene imaging system for monitoring microRNA biogenesis
Hae Young Ko1,2, Do Won Hwang1, Dong Soo Lee1,3 & Soonhag Kim4
Abstract
MicroRNAs (miRNAs), non-coding RNA molecules, have emerged as a part of key gene regulation, participating in a variety of biological processes such as cell development. Current research methods, including northern blot and real-time PCR analysis, have been used to quantify miRNA expression. Major disadvantages of these methods include invasive techniques, such as a tissue biopsy, and the absence of repetitive studies. In this protocol we describe a simple non-invasive imaging method for monitoring miRNAs during neurogenesis. This novel method includes the design of an miRNA reporter gene vector, cell transfection, in vitro luciferase assay and in vivo bioluminescence imaging of miRNAs. Our reporter imaging system allows for repetitive, non-invasive detection of miRNAs, illustrating the miRNA124a (miR124a)-dependent decrease of Gaussia reporter activity during neuronal differentiation. Using this method, construction of a reporter-imaging vector, in vitro and in vivo signal detection steps can be carried out in 
10 d.
- Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.
- Interdisciplinary Course of Radiation Applied Life Science, Seoul National University College of Medicine, Seoul, Korea.
- Department of Nuclear Medicine and Department of Molecular Medicine and Biopharmaceutical Science, College of Medicine, Seoul, Korea.
- Laboratory of Molecular Imaging, Department of Applied Bioscience, CHA Stem Cell Institute, CHA University, Seoul, Korea.
Correspondence to: Soonhag Kim4 e-mail: kimsoonhag@empal.com
Correspondence to: Dong Soo Lee1,3 e-mail: dsl@plaza.snu.ac.kr
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