Isolation of Mitochondria from Liver and Extraction of Total RNA and Protein: Analyses of miRNA and Protein Expression

André L. Simão; Pedro M. Borralho; Clifford J. Steer; Rui E. Castro; Cecília M.P. Rodrigues

doi:10.1007/978-1-0716-1433-4_1

Isolation of Mitochondria from Liver and Extraction of Total RNA and Protein: Analyses of miRNA and Protein Expression

André L. Simão, Pedro M. Borralho, Clifford J. Steer, Rui E. Castro, Cecília M.P. Rodrigues

Medicine - Gastro, Hepatology, Nutrition Division

Research output: Chapter in Book/Report/Conference proceeding › Chapter

4 Scopus citations

Abstract

Several studies have indicated the presence of microRNAs (miRNAs) within mitochondria although the origin, as well as the biological function, of these mitochondrially located miRNAs is largely unknown. The identification and significance of this subcellular localization is gaining increasing relevance to the pathogenesis of certain disease states. Here, we describe the isolation of highly purified mitochondria from rat liver by differential centrifugation, followed by RNAse A treatment to eliminate contaminating RNA. The coupled extraction of total RNA and protein is a more efficient design for allowing the downstream evaluation of miRNA and protein expression in mitochondria.

Original language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	1-15
Number of pages	15
DOIs	https://doi.org/10.1007/978-1-0716-1433-4_1
State	Published - 2021

Publication series

Name	Methods in Molecular Biology
Volume	2310
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Funding Information:
CMPR is supported by the EU H2020 Marie Sklodowska-Curie Project Foie Gras (grant 722619) and by FCT and FEDER (grant PTDC/MEDFAR/29097/2017 e LISBOA-01-0145-FEDER-029097).

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Differential centrifugation
Mitochondria
Protein
TRIzol™
miRNA

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

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abstract = "Several studies have indicated the presence of microRNAs (miRNAs) within mitochondria although the origin, as well as the biological function, of these mitochondrially located miRNAs is largely unknown. The identification and significance of this subcellular localization is gaining increasing relevance to the pathogenesis of certain disease states. Here, we describe the isolation of highly purified mitochondria from rat liver by differential centrifugation, followed by RNAse A treatment to eliminate contaminating RNA. The coupled extraction of total RNA and protein is a more efficient design for allowing the downstream evaluation of miRNA and protein expression in mitochondria.",

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note = "Funding Information: CMPR is supported by the EU H2020 Marie Sklodowska-Curie Project Foie Gras (grant 722619) and by FCT and FEDER (grant PTDC/MEDFAR/29097/2017 e LISBOA-01-0145-FEDER-029097). Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",

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AU - Borralho, Pedro M.

AU - Steer, Clifford J.

AU - Castro, Rui E.

AU - Rodrigues, Cecília M.P.

N1 - Funding Information: CMPR is supported by the EU H2020 Marie Sklodowska-Curie Project Foie Gras (grant 722619) and by FCT and FEDER (grant PTDC/MEDFAR/29097/2017 e LISBOA-01-0145-FEDER-029097). Publisher Copyright: © 2021, Springer Science+Business Media, LLC, part of Springer Nature.

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AB - Several studies have indicated the presence of microRNAs (miRNAs) within mitochondria although the origin, as well as the biological function, of these mitochondrially located miRNAs is largely unknown. The identification and significance of this subcellular localization is gaining increasing relevance to the pathogenesis of certain disease states. Here, we describe the isolation of highly purified mitochondria from rat liver by differential centrifugation, followed by RNAse A treatment to eliminate contaminating RNA. The coupled extraction of total RNA and protein is a more efficient design for allowing the downstream evaluation of miRNA and protein expression in mitochondria.

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Isolation of Mitochondria from Liver and Extraction of Total RNA and Protein: Analyses of miRNA and Protein Expression

Abstract

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