Improving the Precision of Base Editing by Bubble Hairpin Single Guide RNA
Zhiwei Hu, Yannan Wang, Qian Liu, Yan Qiu, Zhiyu Zhong, Kangdi Li, Wenhua Li, Zixin Deng, Yuhui Sun
Abstract
Base editors are DSB-free genome editing tools and have been widely used in diverse living systems. However, it is reported that these tools can cause substantial off-target editings. To meet this challenge, we developed a new approach to improve the specificity of base editors by using hairpin sgRNAs with a bubble. Furthermore, our sgRNA design also dramatically reduced indels and unwanted base substitutions at on-target sites. We believe that the BH-sgRNA design is a significant improvement over existing sgRNAs of base editors, and our design promises to be adaptable to various base editors. We expect that it will make contributions to improving the safety of gene therapy.