Programmable A-to-Y base editing by fusing an adenine base editor with an N-methylpurine DNA glycosylase

Tong, Huawei; Wang, Xuchen; Liu, Yuanhua; Liu, Nana; Li, Yun; Luo, Jianmin; Qian, Ma; Wu, Danni; Li, Jiyong; Xu, Chunhe; Yang, Hui

doi:10.1038/s41587-022-01595-6

Cited by 108 publications

(89 citation statements)

References 20 publications

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“…3a and Supplementary Fig. 6i), consistent with previous findings of gGBE 3 , AYBE 9 and CGBEs [11][12][13][14][15] .…”

Section: Characterization Of Gtbev3 At Human Genomic Dna Sitessupporting

confidence: 92%

“…The deaminase-based base editor (dBE) and deaminase-free glycosylase-based base editor (gBE) are currently two main categories of DNA base editors 3 , enabling direct editing of adenine (A), cytosine (C), or guanine (G), but not thymine (T). In this study, we engineered two orthogonal base editors, gTBE and gCBE, that could achieve highly efficient T and C editing in both cultured human cells and mouse embryos.…”

Section: Discussionmentioning

confidence: 99%

“…Encouraged by the development of gGBE in our previous study 3 , we attempted to develop thymine and cytosine base editor using the deaminase-free glycosylase-based strategy. Since the three pyrimidine bases (i.e., T, C, and U) are structurally similar, we speculated that excision of canonical T or C could be achieved by engineering certain uracil DNA glycosylase, thus triggering the BER pathway and facilitating direct T editing or C editing following the one-step generation of AP sites (Fig.…”

Section: Development Of Orthogonal Base Editors Based On Engineered G...mentioning

confidence: 99%

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Development of deaminase-free T-to-S base editor and C-to-G base editor by engineered human uracil DNA glycosylase

Tong,

Wang,

Liu

et al. 2024

Preprint

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DNA base editors could enable direct editing of adenine (A), cytosine (C), or guanine (G), but there is no base editor for direct thymine (T) editing currently. Here, by fusing Cas9 nickase (nCas9) with engineered human uracil DNA glycosylase (UNG) variants, we developed a deaminase-free glycosylase-based thymine base editor (gTBE) with the ability of direct T editing. By several rounds of UNG mutagenesis via rational screening, we demonstrated that gTBE with engineered UNG variants could achieve T editing efficiency by up to 81.5%. Furthermore, the gTBE exhibited high T-to-S (i.e., T-to-C or T-to-G) conversion ratio with up to 0.97 in cultured human cells. Using similar strategy, we developed a deaminase-free cytosine base editor (gCBE) facilitating specifically direct C editing by engineered UNG with mutations different from gTBE. Thus, we provide two novel base editors, gTBE and gCBE, with corresponding engineered UNG variants, broadening the targeting scope of base editors.

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“…3a and Supplementary Fig. 6i), consistent with previous findings of gGBE 3 , AYBE 9 and CGBEs [11][12][13][14][15] .…”

Section: Characterization Of Gtbev3 At Human Genomic Dna Sitessupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Development Of Orthogonal Base Editors Based On Engineered G...mentioning

confidence: 99%

See 1 more Smart Citation

Development of deaminase-free T-to-S base editor and C-to-G base editor by engineered human uracil DNA glycosylase

Tong,

Wang,

Liu

et al. 2024

Preprint

Self Cite

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“…One such glycosylase base editor is currently being used in a clinical trial by Bioray Laboratories (discussed below). This same strategy was recently applied to ABEs as well, where an engineered hypoxanthine glycosylase enzyme (derived from N-methylpurine DNA glycosylase, MPG) was fused to an ABE, resulting in an adenine transversion base editor (AYBE) that mutagenizes target adenines, with an A•T to C•G base pair as the most common overall outcome [ 17 ].…”

Section: Base Editing Technologiesmentioning

confidence: 99%

In the business of base editors: Evolution from bench to bedside

Porto

Komor

2023

PLoS Biol

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With the advent of recombinant DNA technology in the 1970s, the idea of using gene therapies to treat human genetic diseases captured the interest and imagination of scientists around the world. Years later, enabled largely by the development of CRISPR-based genome editing tools, the field has exploded, with academic labs, startup biotechnology companies, and large pharmaceutical corporations working in concert to develop life-changing therapeutics. In this Essay, we highlight base editing technologies and their development from bench to bedside. Base editing, first reported in 2016, is capable of installing C•G to T•A and A•T to G•C point mutations, while largely circumventing some of the pitfalls of traditional CRISPR/Cas9 gene editing. Despite their youth, these technologies have been widely used by both academic labs and therapeutics-based companies. Here, we provide an overview of the mechanics of base editing and its use in clinical trials.

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“…To further expand the editing competency of the system, dual-function BEs which enable concurrent cytosine and adenine editing were developed by simultaneously fusing cytosine and adenine deaminases into Cas9 variants and applied in mammalian cells and plants ( 82 – 87 ). Most recently, an adenine transversion base editor, AYBE, was developed for A-C and A-T transversion editing in mammalian cells by fusing an ABE with hypoxanthine excision protein N-methylpurine DNA glycosylase (MPG) ( 88 ).…”

Section: Gene Editing Systems Mediate Efficient and Precise Genome Ed...mentioning

confidence: 99%

Advances in CRISPR/Cas gene therapy for inborn errors of immunity

Liu

et al. 2023

Front. Immunol.

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Inborn errors of immunity (IEIs) are a group of inherited disorders caused by mutations in the protein-coding genes involved in innate and/or adaptive immunity. Hematopoietic stem cell transplantation (HSCT) is a mainstay definitive therapy for many severe IEIs. However, the lack of HLA-matched donors increases the risk of developing severe immunological complications. Gene therapy provides long-term clinical benefits and could be an attractive therapeutic strategy for IEIs. In this review, we describe the development and evolution of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated proteins (Cas) gene-editing systems, including double-strand break (DSB)-based gene editing and DSB-free base editing or prime editing systems. Here, we discuss the advances in and issues associated with CRISPR/Cas gene editing tools and their potential as therapeutic alternatives for IEIs. We also highlight the progress of preclinical studies for the treatment of human genetic diseases, including IEIs, using CRISR/Cas and ongoing clinical trials based on this versatile technology.

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Programmable A-to-Y base editing by fusing an adenine base editor with an N-methylpurine DNA glycosylase

Cited by 108 publications

References 20 publications

Development of deaminase-free T-to-S base editor and C-to-G base editor by engineered human uracil DNA glycosylase

Development of deaminase-free T-to-S base editor and C-to-G base editor by engineered human uracil DNA glycosylase

In the business of base editors: Evolution from bench to bedside

Advances in CRISPR/Cas gene therapy for inborn errors of immunity

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