Mol. Cells 2019; 42(): 189
MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates
Songhee H. Kim1,2, Melissa Vieira2,3, Hye-Jin Kim2, Mahipal Singh Kesawat2, and Hye Yoon Park1,2,3,4,*
1Department of Physics and Astronomy, 2Institute of Molecular Biology and Genetics, 3Interdisciplinary Program in Neuroscience, 4Institute of Applied Physics, Seoul National University, Seoul 08826, Korea
Received October 2, 2018; Revised January 17, 2019; Accepted February 5, 2019.; Published online March 28, 2019.
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The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at singlemolecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of β-actin mRNA extracted from the Actb-MBS knock-in and MBS×MCP hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of β-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.
Keywords: β-actin mRNA, mouse, MS2-GFP system, Northern blot, single RNA imaging

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28 February 2019 Volume 42,
Number 2, pp. 97~188

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