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Heterologous Expression and Purification of a CRISPR-Cas9-Based Adenine Base Editor
Methods in Molecular Biology, 2023CRISPR-cas9-guided adenine base editors (ABEs) site-specifically convert the A-T base pair to G-C base pair in genomic DNA. The intracellular delivery of ABE proteins preassembled with guide RNAs (gRNAs) has shown greatly reduced off-target effects compared with that of plasmids or viral vectors containing ABE and gRNA-encoding sequences. For efficient
Seu-Na Lee, Jae-Sung Woo
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Adenine base editors catalyze cytosine conversions in human cells
Nature Biotechnology, 2019 Adenine base editors comprise an adenosine deaminase, evolved in vitro, and a Cas9 nickase. Here, we show that in addition to converting adenine to guanine, adenine base editors also convert cytosine to guanine or thymine in a narrow editing window (positions 5-7) and in a confined TC*N sequence context.
Heon Seok Kim +4 more
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Dual base editor catalyzes both cytosine and adenine base conversions in human cells
Nature Biotechnology, 2020Although base editors are useful tools for precise genome editing, current base editors can only convert either adenines or cytosines. We developed a dual adenine and cytosine base editor (A&C-BEmax) by fusing both deaminases with a Cas9 nickase to achieve C-to-T and A-to-G conversions at the same target site. Compared to single base editors, A&C-BEmax'
Biyun Zhu, Shuming Yin, Li Yang
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Genome-wide target specificity of CRISPR RNA-guided adenine base editors
Nature Biotechnology, 2019 Adenine base editors1 enable efficient targeted adenine-to-guanine single nucleotide conversions to induce or correct point mutations in human cells, animals, and plants1-4. Here we present a modified version of Digenome-seq, an in vitro method for identifying CRISPR (clustered regularly interspaced short palindromic repeats)-induced double-strand ...
Daesik Kim +4 more
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Correcting a patient-specific Rhodopsin mutation with adenine base editor in a mouse model
Molecular TherapyGenome editing offers a great promise to treating human genetic diseases. To assess genome-editing-mediated therapeutic effects in vivo, an animal model is indispensable. The genomic disparities between mice and humans often impede the direct clinical application of genome-editing-mediated treatments using conventional mouse models.
Feng Gu
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Genome Engineering Human ESCs or iPSCs with Cytosine and Adenine Base Editors
2022The ability to engineer specific mutations in human embryonic stem cells (ECSs) or induced pluripotent stem cells (iPSCs) is extremely important in the modeling of human diseases and the study of biological processes. While CRISPR/Cas9 can robustly generate gene knockouts (KOs) and gene loci modifications in coding sequences of iPSCs, it remains ...
Giulia, Pavani +3 more
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An adenine base editor variant expands context compatibility
Nature BiotechnologyAdenine base editors (ABEs) are precise gene-editing agents that convert A:T pairs into G:C through a deoxyinosine intermediate. Existing ABEs function most effectively when the target A is in a TA context. Here we evolve the Escherichia coli transfer RNA-specific adenosine deaminase (TadA) to generate TadA8r, which extends potent deoxyadenosine ...
Yu-Lan Xiao, Yuan Wu, Weixin Tang
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Germline Editing of Drosophila Using CRISPR-Cas9-Based Cytosine and Adenine Base Editors
The CRISPR Journal, 2023Target-AID, BE3, and ABE7.10 base editors fused to the catalytically modified Cas9 and xCas9(3.7) were tested for germline editing of the fruit fly Drosophila melanogaster . We developed a guide RNA-expressing construct, white- 4gRNA, targeting splice ...
Nirav Thakkar +3 more
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