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

Nat Biotechnol.. 2018-05;

Koblan LW, Doman JL, Wilson C, Levy JM, Tay T1, Newby GA, Maianti JP, Raguram A, Liu DR

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Codon Optimization	GenScript codon optimization of bis-bpNLS ABE7.10 (ABEmax) resulted in 1.3- to 7.9-fold higher editing levels than IDT codon usage (in ABE7.10) at high and low plasmid doses, respectively	Get A Quote

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.

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