Abstract
Localization of mRNA is a critical mechanism used by a large fraction of transcripts to restrict its translation to specific cellular regions. Although current high-resolution imaging techniques provide ample information, the analysis methods for localization have either been qualitative or employed quantification in nonrandomly selected regions of interest. Here, we describe an analytical method for objective quantification of mRNA localization using a combination of two characteristics of its molecular distribution, polarization and dispersion. The validity of the method is demonstrated using single-molecule FISH images of budding yeast and fibroblasts. Live-cell analysis of endogenous β-actin mRNA in mouse fibroblasts reveals that mRNA polarization has a half-life of ∼16 min and is cross-correlated with directed cell migration. This novel approach provides insights into the dynamic regulation of mRNA localization and its physiological roles.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 179-184 |
| Number of pages | 6 |
| Journal | Cell reports |
| Volume | 1 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 23 2012 |
| Externally published | Yes |
Bibliographical note
Funding Information:Microscopy equipment for the live cell imaging experiments was provided by the Gruss Lipper Biophotonics Center. This work was supported by NIH GM57071, GM84364 and GM86217 to R.H.S. H.Y.P. was supported by National Research Service Awards F32-GM087122.