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Monitoring the temporal and spatial distribution of RNA in living yeast cells. Methods Mol Biol 2008;419:187-96

Date

03/29/2008

Pubmed ID

18369984

DOI

10.1007/978-1-59745-033-1_13

Scopus ID

2-s2.0-42949178004

Abstract

RNA localization is a cellular process to spatially restrict translation of specific proteins to defined regions within or between cells. Most localized mRNAs contain cis-acting localization elements in the 3'-untranslated region (UTR), which are sufficient for localization of an mRNA to a particular region of the cell. The cis-acting localization elements serve as assembly sites for trans-acting factors which function to sort the mRNA to the correct sub-cellular destination. Although fluorescent in situ hybridization (FISH) has been widely used to study mRNA localization, FISH has a weakness in that it is a static assay, as FISH requires that cells be fixed before hybridization. Consequently, FISH is not ideally suited for investigating dynamic mRNA localization processes. This limitation of FISH has been overcome by the development of techniques that allow the visualization of mRNA in living cells. Here, we present a protocol that tethers green fluorescent protein (GFP) to an mRNA of interest, allowing for the visualization of dynamic mRNA localization processes in living cells.

Author List

Long RM, Urbinati CR

Author

Roy M. Long PhD Assistant Dean, Associate Professor in the Medical School Regional Campuses department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

3' Untranslated Regions
Cloning, Molecular
Green Fluorescent Proteins
Kinetics
Levivirus
Microscopy, Fluorescence
Plasmids
RNA, Fungal
RNA, Messenger
Recombinant Fusion Proteins
Saccharomyces cerevisiae
Subcellular Fractions
Transformation, Genetic
Viral Core Proteins