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Mitoribosome Profiling from Human Cell Culture: A High Resolution View of Mitochondrial Translation

Part of the Methods in Molecular Biology book series (MIMB,volume 2192)

Abstract

Ribosome profiling (Ribo-Seq) is a technique that allows genome-wide, quantitative analysis of translation. In recent years, it has found multiple applications in studies of translation in diverse organisms, tracking protein synthesis with single codon resolution. Traditional protocols applied for generating Ribo-Seq libraries from mammalian cell cultures are not suitable to study mitochondrial translation due to differences between eukaryotic cytosolic and mitochondrial ribosomes. Here, we present an adapted protocol enriching for mitoribosome footprints. In addition, we describe the preparation of small RNA sequencing libraries from the resultant mitochondrial ribosomal protected fragments (mtRPFs).

Key words

  • Mitoribosome
  • Ribosome profiling
  • Mitochondria
  • MitoRibo-Seq

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  • DOI: 10.1007/978-1-0716-0834-0_14
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Correspondence to Joanna Rorbach .

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Pearce, S.F., Cipullo, M., Chung, B., Brierley, I., Rorbach, J. (2021). Mitoribosome Profiling from Human Cell Culture: A High Resolution View of Mitochondrial Translation. In: Minczuk, M., Rorbach, J. (eds) Mitochondrial Gene Expression. Methods in Molecular Biology, vol 2192. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0834-0_14

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  • DOI: https://doi.org/10.1007/978-1-0716-0834-0_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0833-3

  • Online ISBN: 978-1-0716-0834-0

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