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

Research output: Contribution to journalArticlepeer-review

Abstract

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).

Original languageEnglish (US)
Article number101288
JournalSTAR Protocols
Volume3
Issue number2
DOIs
StatePublished - Jun 17 2022

Keywords

  • CRISPR
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Sequencing

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Medicine(all)

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