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KMID : 0578320170400110823
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Molecules and Cells
2017 Volume.40 No. 11 p.823 ~ p.827
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Targeted Base Editing via RNA-Guided Cytidine Deaminases in Xenopus laevis Embryos
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Park Dong-Seok
Yoon Mi-Jung
Kweon Ji-Yeon
Jang An-Hee
Kim Yong-Sub
Choi Sun-Cheol
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Abstract
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Genome editing using programmable nucleases such as CRISPR/Cas9 or Cpf1 has emerged as powerful tools for gene knock-out or knock-in in various organisms. While most genetic diseases are caused by point mutations, these genome-editing approaches are inefficient in inducing single-nucleotide substitutions. Recently, Cas9-linked cytidine deaminases, named base editors (BEs), have been shown to convert cytidine to uridine efficiently, leading to targeted single-base pair substitutions in human cells and organisms. Here, we first report on the generation of Xenopus laevis mutants with targeted single-base pair substitutions using this RNA-guided programmable deaminase. Injection of base editor 3 (BE3) ribonucleoprotein targeting the tyrosinase (tyr) gene in early embryos can induce site-specific base conversions with the rates of up to 20.5%, resulting in oculocutaneous albinism phenotypes without off-target mutations. We further test this base-editing system by targeting the tp53 gene with the result that the expected single-base pair substitutions are observed at the target site. Collectively, these data establish that the programmable deaminases are efficient tools for creating targeted point mutations for human disease modeling in Xenopus.
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KEYWORD
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base editing, CRISPR/Cas9, genome engineering, Xenopus laevis
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