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http://dx.doi.org/10.14348/molcells.2019.2398

MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates  

Kim, Songhee H. (Department of Physics and Astronomy, Seoul National University)
Vieira, Melissa (Institute of Molecular Biology and Genetics, Seoul National University)
Kim, Hye-Jin (Institute of Molecular Biology and Genetics, Seoul National University)
Kesawat, Mahipal Singh (Institute of Molecular Biology and Genetics, Seoul National University)
Park, Hye Yoon (Department of Physics and Astronomy, Seoul National University)
Publication Information
Molecules and Cells / v.42, no.4, 2019 , pp. 356-362 More about this Journal
Abstract
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 single-molecule 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 ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ 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 ${\beta}$-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
${\beta}$-actin mRNA; mouse; MS2-GFP system; Northern blot; single RNA imaging;
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