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 inves-tigate 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.
|Original language||English (US)|
|Number of pages||7|
|Journal||Molecules and cells|
|State||Published - 2019|
Bibliographical noteFunding Information:
This work was supported by the Creative-Pioneering Researchers Program through Seoul National University and the Howard Hughes Medical Institute (HHMI)-Wellcome International Scholar Awards from the Wellcome Trust [208468/Z/17/Z].
© The Korean Society for Molecular and Cellular Biology.
- MS2-GFP system
- Northern blot
- single RNA imaging
- β-actin mRNA