Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction

Luke W Koblan; Jordan L Doman; Christopher Wilson; Jonathan M Levy; Tristan Tay; Gregory A Newby; Juan Pablo Maianti; Aditya Raguram; David R Liu

doi:10.1038/nbt.4172

Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction

Nat Biotechnol. 2018 Oct;36(9):843-846. doi: 10.1038/nbt.4172. Epub 2018 May 29.

Authors

Luke W Koblan^{1

2

3}, Jordan L Doman^{1

2

3}, Christopher Wilson^{1

2

3}, Jonathan M Levy^{1

2

3}, Tristan Tay^{1

2

3}, Gregory A Newby^{1

2

3}, Juan Pablo Maianti^{1

2

3}, Aditya Raguram^{1

2

3}, David R Liu^{1

2

3}

Affiliations

¹ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.
² Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
³ Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.

Abstract

Base editors enable targeted single-nucleotide conversions in genomic DNA. Here we show that expression levels are a bottleneck in base-editing efficiency. We optimize cytidine (BE4) and adenine (ABE7.10) base editors by modification of nuclear localization signals (NLS) and codon usage, and ancestral reconstruction of the deaminase component. The resulting BE4max, AncBE4max, and ABEmax editors correct pathogenic SNPs with substantially increased efficiency in a variety of mammalian cell types.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adenine / metabolism*
CRISPR-Cas Systems
Codon
Cytidine / genetics*
DNA / genetics*
Gene Editing
HEK293 Cells
Humans

Substances

Codon
Cytidine
DNA
Adenine

Abstract

Publication types

MeSH terms

Substances

Grants and funding