作者
B W Thuronyi, Luke W Koblan, Jonathan M Levy, Wei-Hsi Yeh, Christine Zheng, Gregory A Newby, Christopher Wilson, Mantu Bhaumik, Olga Shubina-Oleinik, Jeffrey R Holt, David R Liu
发表日期
2019/9
期刊
Nature biotechnology
卷号
37
期号
9
页码范围
1070-1079
出版商
Nature Publishing Group US
简介
Base editors use DNA-modifying enzymes targeted with a catalytically impaired CRISPR protein to precisely install point mutations. Here, we develop phage-assisted continuous evolution of base editors (BE–PACE) to improve their editing efficiency and target sequence compatibility. We used BE–PACE to evolve cytosine base editors (CBEs) that overcome target sequence context constraints of canonical CBEs. One evolved CBE, evoAPOBEC1-BE4max, is up to 26-fold more efficient at editing cytosine in the GC context, a disfavored context for wild-type APOBEC1 deaminase, while maintaining efficient editing in all other sequence contexts tested. Another evolved deaminase, evoFERNY, is 29% smaller than APOBEC1 and edits efficiently in all tested sequence contexts. We also evolved a CBE based on CDA1 deaminase with much higher editing efficiency at difficult target sites. Finally, we used data from evolved …
学术搜索中的文章
BW Thuronyi, LW Koblan, JM Levy, WH Yeh, C Zheng… - Nature biotechnology, 2019