Protocol for engineering and validating a synthetic mitochondrial intermembrane bridge in mammalian cells

Abstract

Summary: Membrane contact sites are recognized as critical means of intercompartmental communication. Here, we describe a protocol for engineering and validating a synthetic bridge between the inner and outer mitochondrial membranes to support functioning of the endogenous mitochondrial contact site and cristae organizing system (MICOS). A chimeric protein, MitoT, is stably expressed in cultured mammalian cells to bridge the mitochondrial membranes. This approach can be a valuable tool to study the function of the MICOS complex and associated proteins.For complete details on the use and execution of this protocol, please refer to Viana et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Biotechnology and bioengineering
• Cell Biology
• Cell culture
• Cell isolation
• Cell Membrane
• Flow Cytometry/Mass Cytometry