Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage
Alexis C. Komor, Yongjoo B. Kim, Michael S. Packer, John A. Zuris, David R. Liu
Summary
This paper introduced cytosine base editing (CBE), a strategy that fuses a catalytically impaired Cas9 to a cytidine deaminase to directly convert C-G base pairs to T-A in genomic DNA without inducing double-strand breaks or requiring a donor template. The authors showed that within a programmable target window the deaminase converts cytosine to uracil, which is then read as thymine, and that inhibiting base-excision repair markedly improves editing efficiency. The approach achieved precise single-base correction in human and other mammalian cells.
Key findings
- Fused a cytidine deaminase to dCas9/nickase Cas9 to perform direct C-to-T (G-to-A) conversion without double-strand breaks
- Defined a roughly 5-nucleotide activity window within the protospacer for editable cytosines
- Adding a uracil glycosylase inhibitor and using a nickase substantially increased editing purity and efficiency in mammalian cells
Subjects & keywords
Cite this paper
Alexis C. Komor, Yongjoo B. Kim, Michael S. Packer, John A. Zuris, & David R. Liu (2016). Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature. https://doi.org/10.1038/nature17946
@article{komor2016programmable,
author = {Alexis C. Komor and Yongjoo B. Kim and Michael S. Packer and John A. Zuris and David R. Liu},
title = {Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage},
journal = {Nature},
year = {2016},
doi = {10.1038/nature17946},
url = {https://doi.org/10.1038/nature17946}
}