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

• Published in: Molecules and cells Vol. 42; no. 4; pp. 356 - 362
• Main Authors: Kim, Songhee H, Vieira, Melissa, Kim, Hye-Jin, Kesawat, Mahipal Singh, Park, Hye Yoon
• Format: Journal Article
• Language: Korean
• Published: 2019
• Subjects: mouse; MS2-GFP system; Northern blot; single RNA imaging; {\beta}$-actin mRNA
• ISSN: 1016-8478; 0219-1032

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.