Using tRNA-linked molecular beacons to image cytoplasmic mRNAs in live cells

Musa M. Mhlanga, Sanjay Tyagi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Imaging products of gene expression in live cells will provide unique insights into the biology of cells. Molecular beacons make attractive probes for imaging mRNA in live cells as they can report the presence of an RNA target by turning on the fluorescence of a quenched fluorophore. However, when oligonucleotide probes are introduced into cells, they are rapidly sequestered in the nucleus, making the detection of cytoplasmic mRNAs difficult. We have shown that if a molecular beacon is linked to a tRNA, it stays in the cytoplasm and permits detection of cytoplasmic mRNAs. Here we describe two methods of linking molecular beacons to tRNA and show how the joint molecules can be used for imaging an mRNA that is normally present in the cytoplasm in live cultured cells. This protocol should take a total of 4 d to complete.

Original languageEnglish (US)
Pages (from-to)1392-1398
Number of pages7
JournalNature Protocols
Volume1
Issue number3
DOIs
StatePublished - Aug 1 2006

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Transfer RNA
Messenger RNA
Imaging techniques
Cytoplasm
Oligonucleotide Probes
Fluorophores
Gene expression
Cell Biology
Cultured Cells
Joints
Fluorescence
Cells
RNA
Gene Expression
Molecules

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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Using tRNA-linked molecular beacons to image cytoplasmic mRNAs in live cells. / Mhlanga, Musa M.; Tyagi, Sanjay.

In: Nature Protocols, Vol. 1, No. 3, 01.08.2006, p. 1392-1398.

Research output: Contribution to journalArticle

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