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Lim, Kayeong
유전체 교정 연구단
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Nuclear and mitochondrial DNA editing in human cells with zinc finger deaminases

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Title
Nuclear and mitochondrial DNA editing in human cells with zinc finger deaminases
Author(s)
Kayeong Lim; Sung-Ik Cho; Jin-Soo Kim
Publication Date
2022-01-18
Journal
NATURE COMMUNICATIONS, v.13, no.1, pp.1 - 10
Publisher
NATURE PORTFOLIO
Abstract
Base editing in nuclear DNA and mitochondrial DNA (mtDNA) is broadly useful for biomedical research, medicine, and biotechnology. Here the authors present zinc finger deaminases which catalyze targeted C-to-T base conversions without inducing unwanted indels in human cells. Base editing in nuclear DNA and mitochondrial DNA (mtDNA) is broadly useful for biomedical research, medicine, and biotechnology. Here, we present a base editing platform, termed zinc finger deaminases (ZFDs), composed of custom-designed zinc-finger DNA-binding proteins, the split interbacterial toxin deaminase DddA(tox), and a uracil glycosylase inhibitor (UGI), which catalyze targeted C-to-T base conversions without inducing unwanted small insertions and deletions (indels) in human cells. We assemble plasmids encoding ZFDs using publicly available zinc finger resources to achieve base editing at frequencies of up to 60% in nuclear DNA and 30% in mtDNA. Because ZFDs, unlike CRISPR-derived base editors, do not cleave DNA to yield single- or double-strand breaks, no unwanted indels caused by error-prone non-homologous end joining are produced at target sites. Furthermore, recombinant ZFD proteins, expressed in and purified from E. coli, penetrate cultured human cells spontaneously to induce targeted base conversions, demonstrating the proof-of-principle of gene-free gene therapy.
URI
https://pr.ibs.re.kr/handle/8788114/11308
DOI
10.1038/s41467-022-27962-0
ISSN
2041-1723
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > 1. Journal Papers (저널논문)
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