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

doi:10.1016/j.xpro.2022.101288

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

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-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 language	English (US)
Article number	101288
Journal	STAR Protocols
Volume	3
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2022.101288
State	Published - Jun 17 2022

Keywords

CRISPR
Cell Biology
Genetics
Molecular Biology
Sequencing

ASJC Scopus subject areas

General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology
General Neuroscience

Access to Document

10.1016/j.xpro.2022.101288

Cite this

@article{85a0c47400214a3c8f34fd1a50c5e126,

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

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).",

keywords = "CRISPR, Cell Biology, Genetics, Molecular Biology, Sequencing",

author = "Bibekananda Kar and Ankit Sabharwal and Santiago Restrepo-Castillo and Simone, {Brandon W.} and Clark, {Karl J.} and Ekker, {Stephen C.}",

note = "Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = jun,

day = "17",

doi = "10.1016/j.xpro.2022.101288",

language = "English (US)",

volume = "3",

journal = "STAR Protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - An optimized FusX assembly-based technique to introduce mitochondrial TC-to-TT variations in human cell lines

AU - Kar, Bibekananda

AU - Sabharwal, Ankit

AU - Restrepo-Castillo, Santiago

AU - Simone, Brandon W.

AU - Clark, Karl J.

AU - Ekker, Stephen C.

PY - 2022/6/17

Y1 - 2022/6/17

N2 - 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).

AB - 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).

KW - CRISPR

KW - Cell Biology

KW - Genetics

KW - Molecular Biology

KW - Sequencing

UR - http://www.scopus.com/inward/record.url?scp=85128183971&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85128183971&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2022.101288

DO - 10.1016/j.xpro.2022.101288

M3 - Article

C2 - 35496789

AN - SCOPUS:85128183971

SN - 2666-1667

VL - 3

JO - STAR Protocols

JF - STAR Protocols

IS - 2

M1 - 101288

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this