Human cleaving embryos enable efficient mitochondrial base-editing with DdCBE

Wei, Yinghui; Xu, Chunhe; Hu, Feng; Xu, Kui; Li, Zhifang; Hu, Jing; Zhou, Ling; Wu, Yu; Zuo, Zhenrui; Zuo, Erwei; Wen, Li; Yang, Hui; Zhang, Meiling

doi:10.1038/s41421-021-00372-0

Cited by 24 publications

(19 citation statements)

References 13 publications

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“…Where rodent models do not accurately model human mitochondrial diseases, it may soon be possible to generate nonhuman primate models with mtDNA mutations that more closely resemble human conditions. This possibility is supported by recent reports of DdCBE-mediated base editing in 3PN human embryos by two different groups [ 51 , 52 ].…”

Section: Animal Models For Mitochondrial Dysfunction Using Mitochondr...supporting

confidence: 64%

“…Based on this enzyme, Liu lab [ 39 ] developed DddA-derived cytosine base editors (DdCBEs), the first agents capable of precise C-to-T editing in human mtDNA. Since their initial report, DdCBEs have been utilized for targeted mtDNA editing in mice [ 46 ], rats [ 47 ], zebrafish [ 48 , 49 ], plants [ 50 ], and human embryos [ 51 , 52 ], demonstrating their broad potential. In the following sections, we summarized major developments in mitochondrial cytosine base editing thus far, in chronological order.…”

Section: Mitochondrial Base Editingmentioning

confidence: 99%

“…Moreover, a strong correlation between on-target and off-target editing was observed, suggesting that further optimization would be needed in order to minimize off-target editing before clinical use [ 51 ]. A similar approach was taken by Wei et al [ 52 ], successfully introducing mutations at different target sites using the DdCBE platform. They observed that injections at the cleavage stage resulted in higher C-to-T editing compared to injections at the zygote stage [ 52 ].…”

Section: Animal Models For Mitochondrial Dysfunction Using Mitochondr...mentioning

confidence: 99%

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Mitochondrial Base Editing: Recent Advances towards Therapeutic Opportunities

Kar

Restrepo-Castillo

Sabharwal

et al. 2023

IJMS

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Mitochondria are critical organelles that form networks within our cells, generate energy dynamically, contribute to diverse cell and organ function, and produce a variety of critical signaling molecules, such as cortisol. This intracellular microbiome can differ between cells, tissues, and organs. Mitochondria can change with disease, age, and in response to the environment. Single nucleotide variants in the circular genomes of human mitochondrial DNA are associated with many different life-threatening diseases. Mitochondrial DNA base editing tools have established novel disease models and represent a new possibility toward personalized gene therapies for the treatment of mtDNA-based disorders.

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Section: Animal Models For Mitochondrial Dysfunction Using Mitochondr...supporting

confidence: 64%

Section: Mitochondrial Base Editingmentioning

confidence: 99%

Section: Animal Models For Mitochondrial Dysfunction Using Mitochondr...mentioning

confidence: 99%

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Mitochondrial Base Editing: Recent Advances towards Therapeutic Opportunities

Kar

Restrepo-Castillo

Sabharwal

et al. 2023

IJMS

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“…DdCBEs 1 and zinc finger deaminases (ZFDs) 2 -composed of the split interbacterial toxin DddA tox , a uracil glycosylase inhibitor (UGI), and custom-designed DNA-binding TALE arrays and zinc finger proteins, respectively-are highly versatile genome editing tools that enable targeted C-to-T conversions in the nuclear genome and mitochondrial [2][3][4][5][6][7][8][9] and chloroplast DNA [10][11][12] . DddA tox , an enzymatic moiety in the interbacterial toxin DddA derived from Burkholderia cenocepacia, catalyzes cytosine deamination within double-strand DNA (dsDNA) 13 .…”

mentioning

confidence: 99%

Precision mitochondrial DNA editing with high-fidelity DddA-derived base editors

Lee

Baek

et al. 2022

Nat Biotechnol

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Bacterial toxin DddA-derived cytosine base editors (DdCBEs)—composed of split DddAtox (a cytosine deaminase specific to double-stranded DNA), custom-designed TALE (transcription activator-like effector) DNA-binding proteins, and a uracil glycosylase inhibitor—enable mitochondrial DNA (mtDNA) editing in human cells, which may pave the way for therapeutic correction of pathogenic mtDNA mutations in patients. The utility of DdCBEs has been limited by off-target activity, which is probably caused by spontaneous assembly of the split DddAtox deaminase enzyme, independent of DNA-binding interactions. We engineered high-fidelity DddA-derived cytosine base editors (HiFi-DdCBEs) with minimal off-target activity by substituting alanine for amino acid residues at the interface between the split DddAtox halves. The resulting domains cannot form a functional deaminase without binding of their linked TALE proteins at adjacent sites on DNA. Whole mitochondrial genome sequencing shows that, unlike conventional DdCBEs, which induce hundreds of unwanted off-target C-to-T conversions in human mtDNA, HiFi-DdCBEs are highly efficient and precise, avoiding collateral off-target mutations, and as such, they will probably be desirable for therapeutic applications.

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“…A bacterial cytidine deaminase fused with mito-TALEN (DdCBE) was recently established to induce base editing for C > T transition in mtDNA, 26 with successful application in mice, rats, zebrafish, plants, and human embryos. 27 , 28 , 29 , 30 , 31 More recently, small-sized zinc-finger deaminases (ZFDs) were engineered for precise C > T base editing of nuclear and mitochondrial genes, 32 offering several advantages in therapeutic application. The same research team also developed an A > G base editor for mtDNA (TALED, transcription activator-like effector-linked deaminases), providing a broader scope for mtDNA editing.…”

Section: Introductionmentioning

confidence: 99%

Direct evidence of CRISPR-Cas9-mediated mitochondrial genome editing

et al. 2022

The Innovation

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Human cleaving embryos enable efficient mitochondrial base-editing with DdCBE

Cited by 24 publications

References 13 publications

Mitochondrial Base Editing: Recent Advances towards Therapeutic Opportunities

Mitochondrial Base Editing: Recent Advances towards Therapeutic Opportunities

Precision mitochondrial DNA editing with high-fidelity DddA-derived base editors

Direct evidence of CRISPR-Cas9-mediated mitochondrial genome editing

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