Generating stable isolated mitochondrial recipient clones in mammalian cells using MitoPunch mitochondrial transfer

Alexander J. Sercel; Alexander J. Napior; Alexander N. Patananan; Ting-Hsiang Wu; Pei-Yu Chiou; Michael A. Teitell

STAR Protocols (Dec 2021)

Generating stable isolated mitochondrial recipient clones in mammalian cells using MitoPunch mitochondrial transfer

Alexander J. Sercel,
Alexander J. Napior,
Alexander N. Patananan,
Ting-Hsiang Wu,
Pei-Yu Chiou,
Michael A. Teitell

Affiliations

Alexander J. Sercel: Molecular Biology Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, CA 90095, USA
Alexander J. Napior: Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
Alexander N. Patananan: Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
Ting-Hsiang Wu: NanoCav, LLC, Culver City, CA 90230, USA; NantWorks, LLC, Culver City, CA 90230, USA
Pei-Yu Chiou: Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA
Michael A. Teitell: Molecular Biology Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90024, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 4
p. 100850

Abstract

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Summary: This protocol describes the assembly and use of MitoPunch to deliver mitochondria containing mitochondrial DNA (mtDNA) into cells lacking mtDNA (ρ0 cells). MitoPunch generates stable isolated mitochondrial recipient clones with restored mtDNA and recovered respiration, enabling investigation of mtDNA mutations and mtDNA-nuclear DNA interactions in a range of cell types.For complete details on the use and execution of this protocol, please refer to Sercel et al. (2021) and Patananan et al. (2020).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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