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

doi:10.1016/j.xpro.2021.100850

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

STAR Protoc. 2021 Sep 28;2(4):100850. doi: 10.1016/j.xpro.2021.100850. eCollection 2021 Dec 17.

Authors

Alexander J Sercel¹, Alexander J Napior², Alexander N Patananan², Ting-Hsiang Wu^{3

4}, Pei-Yu Chiou^{5

6

7}, Michael A Teitell^{1

2

6

7

8

9

10}

Affiliations

¹ Molecular Biology Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
³ NanoCav, LLC, Culver City, CA 90230, USA.
⁴ NantWorks, LLC, Culver City, CA 90230, USA.
⁵ Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁶ California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁷ Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁸ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁹ Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
¹⁰ Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90024, USA.

Abstract

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).

Keywords: Biotechnology and bioengineering; Cell Biology; Cell culture; Cell-based Assays; Metabolism.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cells, Cultured
Clone Cells / metabolism
DNA, Mitochondrial* / genetics
Mammals / genetics
Mitochondria* / genetics

Substances

DNA, Mitochondrial

Abstract

Publication types

MeSH terms

Substances

Grants and funding