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Methods. 2019 Mar 2. pii: S1046-2023(18)30318-9. doi: 10.1016/j.ymeth.2019.02.022. [Epub ahead of print]

Base editing the mammalian genome.

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Sandra and Edward Meyer Cancer Center, United States; Weill Cornell/Rockefeller/Sloan Kettering Tri-I MD-PhD Program, New York 10065, United States.
Sandra and Edward Meyer Cancer Center, United States; Department of Medicine, United States.
Sandra and Edward Meyer Cancer Center, United States; Department of Medicine, United States; Department of Biochemistry, Weill Cornell Medicine, New York 10021, United States. Electronic address:


Base editing is a powerful technology that enables programmable conversion of single nucleotides in the mammalian genome. Base editors consist of a partially active Cas9 nuclease (Cas9D10A) tethered to a natural or synthetic DNA modifying enzyme. Though only recently described, BE has already shown enormous potential for basic and translational research, allowing the creation or repair of disease alleles in a variety of cell types and model organisms. In the past 2 years, a vast array of new and modified base editor variants have been described, expanding the flexibility and usefulness of the approach. Though simple in concept, effective implementation of base editing requires an understanding of the advantages and limitations of each of these tools. Here, we provide an overview of the concepts of DNA base editing, and discuss the recent progress toward the development of optimized base editing systems for mammalian cells. In addition, we highlight key technical aspects of designing and executing BE experiments, and provide detailed experimental examples of successful base editing in cell lines and organoids to help guide the effective use of these tools for genome modification.


APOBEC; BE; Base editing; CRISPR

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