A genomic integration method to visualize localization of endogenous mRNAs in living yeast

Liora Haim; Gadi Zipor; Stella Aronov; Jeffrey E. Gerst

doi:10.1038/nmeth1040

A genomic integration method to visualize localization of endogenous mRNAs in living yeast

Liora Haim, Gadi Zipor, Stella Aronov, Jeffrey E. Gerst

Research output: Contribution to journal › Article › peer-review

99 Scopus citations

Abstract

mRNA localization may be an important determinant for protein localization. We describe a simple PCR-based genomic-tagging strategy (m-TAG) that uses homologous recombination to insert binding sites for the RNA-binding MS2 coat protein (MS2-CP) between the coding region and 3′ untranslated region (UTR) of any yeast gene. Upon coexpression of MS2-CP fused with GFP, we demonstrate the localization of endogenous mRNAs (ASH1, SRO7, PEX3 and OXA1) in living yeast (Saccharomyces cerevisiae).

Original language	English
Pages (from-to)	409-412
Number of pages	4
Journal	Nature Methods
Volume	4
Issue number	5
DOIs	https://doi.org/10.1038/nmeth1040
State	Published - May 2007
Externally published	Yes

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title = "A genomic integration method to visualize localization of endogenous mRNAs in living yeast",

abstract = "mRNA localization may be an important determinant for protein localization. We describe a simple PCR-based genomic-tagging strategy (m-TAG) that uses homologous recombination to insert binding sites for the RNA-binding MS2 coat protein (MS2-CP) between the coding region and 3′ untranslated region (UTR) of any yeast gene. Upon coexpression of MS2-CP fused with GFP, we demonstrate the localization of endogenous mRNAs (ASH1, SRO7, PEX3 and OXA1) in living yeast (Saccharomyces cerevisiae).",

author = "Liora Haim and Gadi Zipor and Stella Aronov and Gerst, \{Jeffrey E.\}",

note = "Funding Information: We thank K. Bloom, P. Brennwald, J. Brickner, M. Longtine, S. Michaelis, R. Singer and D. Stillman for the generous gifts of reagents; A. Cohen and I. Goldshtein for technical assistance. This work was generously supported by a grant from the Kahn Fund for Systems Biology, Weizmann Institute of Science. J.E.G. holds the Henry Kaplan Chair in Cancer Research.",

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AU - Zipor, Gadi

AU - Aronov, Stella

AU - Gerst, Jeffrey E.

N1 - Funding Information: We thank K. Bloom, P. Brennwald, J. Brickner, M. Longtine, S. Michaelis, R. Singer and D. Stillman for the generous gifts of reagents; A. Cohen and I. Goldshtein for technical assistance. This work was generously supported by a grant from the Kahn Fund for Systems Biology, Weizmann Institute of Science. J.E.G. holds the Henry Kaplan Chair in Cancer Research.

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N2 - mRNA localization may be an important determinant for protein localization. We describe a simple PCR-based genomic-tagging strategy (m-TAG) that uses homologous recombination to insert binding sites for the RNA-binding MS2 coat protein (MS2-CP) between the coding region and 3′ untranslated region (UTR) of any yeast gene. Upon coexpression of MS2-CP fused with GFP, we demonstrate the localization of endogenous mRNAs (ASH1, SRO7, PEX3 and OXA1) in living yeast (Saccharomyces cerevisiae).

AB - mRNA localization may be an important determinant for protein localization. We describe a simple PCR-based genomic-tagging strategy (m-TAG) that uses homologous recombination to insert binding sites for the RNA-binding MS2 coat protein (MS2-CP) between the coding region and 3′ untranslated region (UTR) of any yeast gene. Upon coexpression of MS2-CP fused with GFP, we demonstrate the localization of endogenous mRNAs (ASH1, SRO7, PEX3 and OXA1) in living yeast (Saccharomyces cerevisiae).

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A genomic integration method to visualize localization of endogenous mRNAs in living yeast

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