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An Unbiased Analysis Method to Quantify mRNA Localization Reveals Its Correlation with Cell Motility
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 employe...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4079260/ https://www.ncbi.nlm.nih.gov/pubmed/22832165 http://dx.doi.org/10.1016/j.celrep.2011.12.009 |
Sumario: | 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 non-randomly 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. |
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