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Import of Fluorescent RNA into Mitochondria of Living Cells

  • Jaroslav Zelenka
  • Petr JežekEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1351)

Abstract

Methods of in vivo visualization and manipulation of mitochondrial genetic machinery are limited due to the need to surpass not only the cytoplasmic membrane but also two mitochondrial membranes. Here, we employ the matrix-addressing sequence of mitochondrial ribosomal 5S-rRNA (termed MAM), which is naturally imported into mammalian mitochondria, to construct an import system for in vivo targeting of mitochondrial (mt) DNA or mtRNA, in order to provide fluorescence hybridization of the desired sequences.

Key words

5S-rRNA Mitochondrial DNA Nucleic acid import into mitochondria Fluorescent in vivo hybridization of mtDNA Mitochondrial nucleoids 

Notes

Acknowledgement

This work was supported by the Czech Science Foundation grant No. P305/12/P388 to JZ and by the research projects RVO67985823 and BIOCEV—Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (CZ.1.05/1.1.00/02.0109), the European Regional Development Fund.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department No. 75, Membrane Transprot BiophysicsInstitute of Physiology, Academy of Sciences of the Czech RepublicPrague 4Czech Republic

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