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

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 ancestr...

Full description

Saved in:
Bibliographic Details
Published in:Nature biotechnology 2018-10, Vol.36 (9), p.843-846
Main Authors: Koblan, Luke W, Doman, Jordan L, Wilson, Christopher, Levy, Jonathan M, Tay, Tristan, Newby, Gregory A, Maianti, Juan Pablo, Raguram, Aditya, Liu, David R
Format: Article
Language:English
Subjects:
Adenine
Adenine - metabolism
Cells (Biology)
Codon
Codons
CRISPR-Cas Systems
Cytidine - genetics
Deoxyribonucleic acid
DNA
DNA - genetics
Editing
Editors
Efficiency
Gene Editing
Gene expression
Gene loci
Gene mutation
Genetic engineering
HEK293 Cells
Humans
Localization
Mammals
Methods
Mutation
Nucleobases
Nucleosides
Nucleotides
Optimization
Optimization theory
Proteins
Purines
Reconstruction
Single-nucleotide polymorphism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt.4172