Multiplex precise base editing in cynomolgus monkeys

Zhang, Wenhui; Aida, Tomomi; Rosario, Ricardo C.H. del; Wilde, Jonathan J.; Ding, Chenhui; Zhang, Xiaohui; Baloch, Zulqarnain; Huang, Yan; Tang, Yu; Li, Duanduan; Lu, Hongsheng; Zhou, Yang; Jiang, Minqing; Xu, Dongdong; Fang, Zhihao; Zheng, Zhan-Hong; Huang, Qunshan; Feng, Guoping; Yang, Shihua

doi:10.1038/s41467-020-16173-0

Cited by 33 publications

(32 citation statements)

References 50 publications

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“…Nonetheless, we observed indels in only 2/60 Dnmt1 embryos and 2/61 Chd2 embryos, demonstrating the usefulness of PE3b for eliminating indels associated with PE3 ( Figure 5G). Therefore, we conclude that PE3b is a viable option for reducing or eliminating indels associated with prime editing but note that its design restrictions, which limit the number of mutations for which it can be applied, necessitate additional advances to improve prime editing for applications that require high efficiency and allele frequency (Aida and Feng, 2020;Zhang et al, 2020;Zhou et al, 2019).…”

Section: Prime Editing By Pe3b In Mouse Embryosmentioning

confidence: 94%

Prime editing primarily induces undesired outcomes in mice

Aida

Wilde

Yang

et al. 2020

Preprint

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SummaryGenome editing has transformed biomedical science, but is still unpredictable and often induces undesired outcomes. Prime editing (PE) is a promising new approach due to its proposed flexibility and ability to avoid unwanted indels. Here, we show highly efficient PE-mediated genome editing in mammalian zygotes. Utilizing chemically modified guideRNAs, PE efficiently introduced 10 targeted modifications including substitutions, deletions, and insertions across 6 genes in mouse embryos. However, we unexpectedly observed a high frequency of undesired outcomes such as large deletions and found that these occurred more often than pure intended edits across all of the edits/genes. We show that undesired outcomes result from the double-nicking PE3 strategy, but that omission of the second nick largely ablates PE function. However, sequential double-nicking with PE3b, which is only applicable to a fraction of edits, eliminated undesired outcomes. Overall, our findings demonstrate the promising potential of PE for predictable, flexible, and highly efficient in vivo genome editing, but highlight the need for improved variations of PE before it is ready for widespread use.

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Section: Prime Editing By Pe3b In Mouse Embryosmentioning

confidence: 94%

Prime editing primarily induces undesired outcomes in mice

Aida

Wilde

Yang

et al. 2020

Preprint

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“…Zhou et al verified the efficiency of the BE system at the embryonic level in mice and macaques, providing an effective reference for the later establishment of T158M mutant RTT animal models [ 62 ]. Zhang et al coinjected SpCas9-based ABE mRNAs, SaCas9-based SaKKH-BE3 mRNAs, and their corresponding sgRNAs into monkey embryos and found that CBE and ABE can function in the same cell, suggesting an effective treatment strategy for polygenic diseases [ 63 ]. The NHP models will further improve our understanding of diseases and help us to comprehensively and accurately evaluate the safety and effectiveness of the drug before the clinic.…”

Section: Development Of the Nhp Modelsmentioning

confidence: 99%

Embryo-Engineered Nonhuman Primate Models: Progress and Gap to Translational Medicine

2021

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Animal models of human diseases are vital in better understanding the mechanism of pathogenesis and essential for evaluating and validating potential therapeutic interventions. As close relatives of humans, nonhuman primates (NHPs) play an increasingly indispensable role in advancing translational medicine research. In this review, we summarized the progress of NHP models generated by embryo engineering, analyzed their unique advantages in mimicking clinical patients, and discussed the remaining gap between basic research of NHP models to translational medicine.

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“…While this review discusses base editing in mouse and human cells, base editing has been successfully applied in plants [ 8 , 9 ] — including cotton [ 10 ], rice [ 11 ], soybean [ 12 ] — zebrafish [ 13 , 14 ], pigs [ 15 , 16 ], rats [ 17 ], rabbits [ 18–20 ], sheep [ 21 , 22 ], and more recently monkeys [ 23 , 24 ].…”

Section: Introductionmentioning

confidence: 99%

CRISPR base editing applications for identifying cancer-driving mutations

Pál

Herold

2021

Biochemical Society Transactions

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CRISPR base editing technology is a promising genome editing tool as (i) it does not require a DNA template to introduce mutations and (ii) it avoids creating DNA double-strand breaks, which can lead to unintended chromosomal alterations or elicit an unwanted DNA damage response. Given many cancers originate from point mutations in cancer-driving genes, the application of base editing for either modelling tumour development, therapeutic editing, or functional screening is of great promise. In this review, we summarise current DNA base editing technologies and will discuss recent advancements and existing hurdles for its usage in cancer research.

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Multiplex precise base editing in cynomolgus monkeys

Cited by 33 publications

References 50 publications

Prime editing primarily induces undesired outcomes in mice

Prime editing primarily induces undesired outcomes in mice

Embryo-Engineered Nonhuman Primate Models: Progress and Gap to Translational Medicine

CRISPR base editing applications for identifying cancer-driving mutations

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