This chapter presents the MS2-GFP system, a method to study the trafficking of RNA molecules in living cells. This system is based on two components: a fusion of the MS2 coat protein to a fluorescent protein and a reporter mRNA containing multimers of the RNA stem-loop recognized by the MS2 coat protein. The MS2-GFP protein bound to the RNA stem-loops acts as a beacon that allows the detection of this mRNA within a cell by epifluorescence or confocal microscopy. This chapter focuses on the use of this system in mammalian fibroblast cells and in yeast cells, and discusses several technical considerations of the MS2-GFP system. Detailed protocols for validating the MS2-GFP signal in fixed cells by fluorescent in situ hybridization of the target RNA using fluorophore-labeled oligonucleotide probes are also provided.
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| Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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| Site | Modification | Modifier | Source | Reference |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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| Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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| Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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