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Updated: Oct 17, 2025
Author Spotlight: Transmitochondrial Cybrid Generation Using Cancer Cell Lines
Published on: March 17, 2023
Generating stable isolated mitochondrial recipient clones in mammalian cells using MitoPunch mitochondrial transfer
Alexander J Sercel1, Alexander J Napior2, Alexander N Patananan2
1Molecular Biology Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
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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).
