Protocol for rapid manipulation of mitochondrial morphology in living cells using inducible counter mitochondrial morphology (iCMM)

Takafumi Miyamoto; Song-Iee Han; Hitoshi Shimano

doi:10.1016/j.xpro.2021.100721

Protocol for rapid manipulation of mitochondrial morphology in living cells using inducible counter mitochondrial morphology (iCMM)

STAR Protoc. 2021 Aug 5;2(3):100721. doi: 10.1016/j.xpro.2021.100721. eCollection 2021 Sep 17.

Authors

Takafumi Miyamoto^{1

2}, Song-Iee Han¹, Hitoshi Shimano¹

Affiliations

¹ Department of Internal Medicine (Endocrinology and Metabolism), Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
² Transborder Medical Research Center, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan.

Abstract

Disruption of mitochondrial morphology occurs during various diseases, but the biological significance is not entirely clear. Here, we describe a detailed step-by-step protocol for a chemically inducible dimerization system-based synthetic protein device, termed inducible counter mitochondrial morphology. This system allows artificial manipulation of mitochondrial morphology on a timescale of minutes in living mammalian cells. We also describe an AI-assisted imaging processing approach. For complete details on the use and execution of this protocol, please refer to Miyamoto et al., 2021.

Keywords: Biotechnology and bioengineering; Cell Biology; Molecular/Chemical Probes; Protein Biochemistry.

Publication types

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

MeSH terms

Dimerization
Genetic Engineering / methods*
HeLa Cells
Humans
Mitochondria / metabolism
Mitochondria / physiology*
Organelle Shape / drug effects*