Sandra and Edward Meyer Cancer Center, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.
Weill Cornell/Rockefeller/Sloan-Kettering Tri-Institutional MD-PhD program, New York, New York, USA.
Nat Biotechnol. 2018 Oct;36(9):888-893. doi: 10.1038/nbt.4194. Epub 2018 Jul 3.
CRISPR base editing enables the creation of targeted single-base conversions without generating double-stranded breaks. However, the efficiency of current base editors is very low in many cell types. We reengineered the sequences of BE3, BE4Gam, and xBE3 by codon optimization and incorporation of additional nuclear-localization sequences. Our collection of optimized constitutive and inducible base-editing vector systems dramatically improves the efficiency by which single-nucleotide variants can be created. The reengineered base editors enable target modification in a wide range of mouse and human cell lines, and intestinal organoids. We also show that the optimized base editors mediate efficient in vivo somatic editing in the liver in adult mice.
CRISPR 碱基编辑可在不产生双链断裂的情况下实现靶向单碱基转换。然而，目前的碱基编辑器在许多细胞类型中的效率非常低。我们通过密码子优化和额外核定位序列的掺入，重新设计了 BE3、BE4Gam 和 xBE3 的序列。我们优化的组成型和诱导型碱基编辑载体系统显著提高了创建单核苷酸变体的效率。经重新设计的碱基编辑器可在广泛的小鼠和人类细胞系以及肠道类器官中实现靶标修饰。我们还表明，优化后的碱基编辑器可介导成年小鼠肝脏中的高效体内体细胞编辑。
