An optimized FusX assembly-based technique to introduce mitochondrial TC-to-TT variations in human cell lines

Bibekananda Kar; Ankit Sabharwal; Santiago Restrepo-Castillo; Brandon W. Simone; Karl J. Clark; Stephen C. Ekker

STAR Protocols (Jun 2022)

An optimized FusX assembly-based technique to introduce mitochondrial TC-to-TT variations in human cell lines

Bibekananda Kar,
Ankit Sabharwal,
Santiago Restrepo-Castillo,
Brandon W. Simone,
Karl J. Clark,
Stephen C. Ekker

Affiliations

Bibekananda Kar: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA
Ankit Sabharwal: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA
Santiago Restrepo-Castillo: Mayo Clinic Graduate School of Biomedical Sciences, Virology and Gene Therapy Track, Mayo Clinic, Rochester, MN, USA
Brandon W. Simone: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA
Karl J. Clark: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA
Stephen C. Ekker: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101288

Abstract

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Summary: The FusX TALE Based Editor (FusXTBE) is a programmable base editing platform that can introduce specific TC-to-TT variations in the mitochondrial DNA (mtDNA). Here, we provide a protocol describing the synthesis and testing of the FusXTBE plasmids in cultured human cell lines. This tool is designed to be easily modified to work in diverse applications where editing of mitochondrial DNA is desired.For complete details on the use and execution of this protocol, please refer to Sabharwal et al. (2021) and Ma et al. (2016).

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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