DdCBE-mediated mitochondrial base editing in human 3PN embryos

Chen, Xiaoxu; Liang, Dong; Guo, Jiayin; Zhang, Junqiang; Sun, Haifeng; Zhang, Xiaolan; Jin, Jiachuan; Dai, Yichen; Bao, Qinmin; Qian, Xiangchen; Tan, Lei; Hu, Ping; Ling, Xiufeng; Shen, Bin; Xu, Zhengfeng

doi:10.1038/s41421-021-00358-y

Cited by 28 publications

(26 citation statements)

References 10 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%

“…In particular, Chen et al [ 51 ] used DdCBE-encoding mRNAs to introduce the m.3733G>A, m.8363G>A, and m.13513G>A mutations in 3PN human embryos, associated with LHON, MERRF, and MELAS or Leigh syndrome, respectively. High levels of on-target base editing for the m.3733G>A and m.8363G>A sites were observed, although editing at the m.13513G>A site was less successful [ 51 ]. As in the in vitro studies, bystander editing was detected around the target sites during testing in human 3PN embryos [ 51 ].…”

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

confidence: 99%

“…High levels of on-target base editing for the m.3733G>A and m.8363G>A sites were observed, although editing at the m.13513G>A site was less successful [ 51 ]. As in the in vitro studies, bystander editing was detected around the target sites during testing in human 3PN embryos [ 51 ]. 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 ].…”

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%

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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“…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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Enhanced C‐To‐T and A‐To‐G Base Editing in Mitochondrial DNA with Engineered DdCBE and TALED

Wei,

Jin,

Huang

et al. 2023

Advanced Science

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Mitochondrial base editing with DddA‐derived cytosine base editor (DdCBE) is limited in the accessible target sequences and modest activity. Here, the optimized DdCBE tools is presented with improved editing activity and expanded C‐to‐T targeting scope by fusing DddA11 variant with different cytosine deaminases with single‐strand DNA activity. Compared to previous DdCBE based on DddA11 variant alone, fusion of the activation‐induced cytidine deaminase (AID) from Xenopus laevis not only permits cytosine editing of 5′‐GC‐3′ sequence, but also elevates editing efficiency at 5′‐TC‐3′, 5′‐CC‐3′, and 5′‐GC‐3′ targets by up to 25‐, 10‐, and 6‐fold, respectively. Furthermore, the A‐to‐G editing efficiency is significantly improved by fusing the evolved DddA6 variant with TALE‐linked deoxyadenosine deaminase (TALED). Notably, the authors introduce the reported high‐fidelity mutations in DddA and add nuclear export signal (NES) sequences in DdCBE and TALED to reduce off‐target editing in the nuclear and mitochondrial genome while improving on‐target editing efficiency in mitochondrial DNA (mtDNA). Finally, these engineered mitochondrial base editors are shown to be efficient in installing mtDNA mutations in human cells or mouse embryos for disease modeling. Collectively, the study shows broad implications for the basic study and therapeutic applications of optimized DdCBE and TALED.

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DdCBE-mediated mitochondrial base editing in human 3PN embryos

Cited by 28 publications

References 10 publications

Mitochondrial Base Editing: Recent Advances towards Therapeutic Opportunities

Mitochondrial Base Editing: Recent Advances towards Therapeutic Opportunities

Direct evidence of CRISPR-Cas9-mediated mitochondrial genome editing

Enhanced C‐To‐T and A‐To‐G Base Editing in Mitochondrial DNA with Engineered DdCBE and TALED

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