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KMID : 0578320190420040356
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Molecules and Cells
2019 Volume.42 No. 4 p.356 ~ p.362
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MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates
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Kim Song-Hee H.
Vieira Melissa
Kim Hye-Jin
Kesawat Mahipal Singh
Park Hye-Yoon
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Abstract
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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 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 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.
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KEYWORD
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β-actin mRNA, mouse, MS2-GFP system, Northern blot, single RNA imaging
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