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Precise mutagenesis in zebrafish using cytosine base editors
Nature Protocols, 2023Base editing is a powerful CRISPR-based technology for introducing precise substitutions into the genome. This technology greatly advances mutagenesis possibilities in vivo, particularly in zebrafish, for which the generation of precise point mutations is still challenging.
Marion Rosello +3 more
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A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing
Nature Biotechnology, 2020 Existing adenine and cytosine base editors induce only a single type of modification, limiting the range of DNA alterations that can be created. Here we describe a CRISPR-Cas9-based synchronous programmable adenine and cytosine editor (SPACE) that can concurrently introduce A-to-G and C-to-T substitutions with minimal RNA off-target edits.
Julian Grünewald +2 more
exaly +5 more sources
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'
Geng Chen +2 more
exaly +3 more sources
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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Adenine base editors catalyze cytosine conversions in human cells
Nature Biotechnology, 2019Adenine 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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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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Precision genome editing using cytosine and adenine base editors in mammalian cells
Nature Protocols, 2021Genome editing has transformed the life sciences and has exciting prospects for use in treating genetic diseases. Our laboratory developed base editing to enable precise and efficient genome editing while minimizing undesired byproducts and toxicity associated with double-stranded DNA breaks.
Tony P. Huang +2 more
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Cytosine base editors go off-target
Nature Reviews Genetics, 2019Two studies in Science show that cytosine base editors, but not adenine base editors or CRISPR–Cas9, induce notable off-target single-nucleotide variants in rice and in mouse embryos.
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Targeted, random mutagenesis of plant genes with dual cytosine and adenine base editors
Nature Biotechnology, 2020Targeted saturation mutagenesis of crop genes could be applied to produce genetic variants with improved agronomic performance. However, tools for directed evolution of plant genes, such as error-prone PCR or DNA shuffling, are limited1. We engineered five saturated targeted endogenous mutagenesis editors (STEMEs) that can generate de novo mutations ...
Chao Li +9 more
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Cas9‐orthologue‐mediated cytosine and adenine base editors recognizing NNAAAA PAM sequences
Biotechnology Journal, 2023AbstractCRISPR/Cas9 system has been applied as an effective genome‐targeting technology. By fusing deaminases with Cas9 nickase (nCas9), various cytosine and adenine base editors (CBEs and ABEs) have been successfully developed that can efficiently induce nucleotide conversions and install pathogenic single nucleotide variants (SNVs) in cultured cells ...
Min Li +8 more
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