Improving the Precision of Base Editing by Bubble Hairpin Single Guide RNA.
Improving the Precision of Base Editing by Bubble Hairpin Single Guide RNA.
复制标题
气泡发夹单向导RNA提高碱基编辑精度
作者：
Hu Z;Wang Y;Liu Q;Qiu Y;Zhong Z;Li K;Li W;Deng Z;Sun Y
碱基编辑器是一种无双链断裂（DSB）的基因组编辑工具，已广泛应用于各种生命系统中。然而，据报道，这些工具可能导致大量的脱靶编辑。碱基编辑是一种强大的基因组编辑方法，可以在没有双链DNA断裂（DSBs）的情况下实现单核苷酸的变化。然而，脱靶效应以及其他不希望在目标位点上的编辑仍然是其应用的障碍。在此，我们报道了气泡发夹单导rna (bubble hairpin single guide rna, bhsgrnas)，它包含一个在5 '端有气泡区域的发夹结构，可以有效地应用于胞嘧啶碱基编辑器（CBE）和腺嘌呤碱基编辑器（ABE），在不牺牲靶上编辑效率的情况下显著减少脱靶编辑。同时，这种设计也提高了碱基编辑器3 （BE3）在靶位点诱导的C-to-T转化的纯度。我们的研究结果提出了一种独特而有效的策略来提高碱基编辑的特异性。
Base editors are double-strand-break (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 editing. Base editing is a powerful genome editing approach that enables single-nucleotide changes without double-stranded DNA breaks (DSBs). However, off-target effects as well as other undesired editings at on-target sites remain obstacles for its application. Here, we report that bubble hairpin single guide RNAs (BH-sgRNAs), which contain a hairpin structure with a bubble region on the 5′ end of the guide sequence, can be efficiently applied to both cytosine base editor (CBE) and adenine base editor (ABE) and significantly decrease off-target editing without sacrificing on-target editing efficiency. Meanwhile, such a design also improves the purity of C-to-T conversions induced by base editor 3 (BE3) at on-target sites. Our results present a distinctive and effective strategy to improve the specificity of base editing.