高效的兔 RNA 引导碱基编辑。

Highly efficient RNA-guided base editing in rabbit.

机构信息

Jilin Provincial Key Laboratory of Animal Embryo Engineering, Institute of Zoonosis, Jilin University, Changchun, 130062, China.

Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong, 510530, China.

出版信息

Nat Commun. 2018 Jul 13;9(1):2717. doi: 10.1038/s41467-018-05232-2.

Cytidine base editors (CBEs) and adenine base editors (ABEs), composed of a cytidine deaminase or an evolved adenine deaminase fused to Cas9 nickase, enable the conversion of C·G to T·A or A·T to G·C base pair in organisms, respectively. Here, we show that BE3 and ABE7.10 systems can achieve a targeted mutation efficiency of 53-88% and 44-100%, respectively, in both blastocysts and Founder (F0) rabbits. Meanwhile, this strategy can be used to precisely mimic human pathologies by efficiently inducing nonsense or missense mutations as well as RNA mis-splicing in rabbit. In addition, the reduced frequencies of indels with higher product purity are also determined in rabbit blastocysts by BE4-Gam, which is an updated version of the BE3 system. Collectively, this work provides a simple and efficient method for targeted point mutations and generation of disease models in rabbit.

胞嘧啶碱基编辑器（CBEs）和腺嘌呤碱基编辑器（ABEs）由胞嘧啶脱氨酶或进化的腺嘌呤脱氨酶与 Cas9 切口酶融合而成，分别能够在生物体中将 C·G 转换为 T·A 或 A·T 转换为 G·C 碱基对。在这里，我们表明 BE3 和 ABE7.10 系统能够分别在囊胚和 Founder（F0）兔中实现 53-88%和 44-100%的靶向突变效率。同时，该策略还可以通过高效诱导无义或错义突变以及 RNA 错剪接，在兔中精确模拟人类疾病。此外，通过 BE4-Gam（BE3 系统的更新版本），在兔囊胚中也确定了具有更高产物纯度的 indels 的频率降低。总的来说，这项工作为兔中靶向点突变和疾病模型的产生提供了一种简单而高效的方法。