Single C-to-T substitution using engineered APOBEC3G-nCas9 base editors with minimum genome- and transcriptome-wide off-target effects

文献类型： 外文期刊

第一作者： Lee, Sangsin

作者： Lee, Sangsin;Li, Jing;Hilton, Isaac B.;Gao, Xue;Ding, Ning;Yuan, Qichen;Yang, Jie;Kolomeisky, Anatoly B.;Gao, Xue;Sun, Yidi;Sun, Yidi;Yuan, Tanglong;Zuo, Erwei;Liu, Lizhong;Hilton, Isaac B.;Wang, Qian;Kolomeisky, Anatoly B.;Kolomeisky, Anatoly B.

作者机构：

期刊名称：SCIENCE ADVANCES （ 影响因子：14.136； 五年影响因子：16.446 ）

ISSN： 2375-2548

年卷期： 2020 年 6 卷 29 期

页码：

收录情况： SCI

摘要： Cytosine base editors (CBEs) enable efficient cytidine-to-thymidine (C-to-T) substitutions at targeted loci without double-stranded breaks. However, current CBEs edit all Cs within their activity windows, generating undesired bystander mutations. In the most challenging circumstance, when a bystander C is adjacent to the targeted c, existing base editors fail to discriminate them and edit both Cs.To improve the precision of CBE, we identified and engineered the human APOBEC3G (A3G) deaminase; when fused to the Cas9 nickase, the resulting A3G-BEs exhibit selective editing of the second C in the 5'-CC-3' motif in human cells. Our A3G-BEs could install a single disease-associated C-to-T substitution with high precision. The percentage of perfectly modified alleles is more than 6000-fold for disease correction and more than 600-fold for disease modeling compared with BE4max. On the basis of the two-cell embryo injection method and RNA sequencing analysis, our A3G-BEs showed minimum genome- and transcriptome-wide off-target effects, achieving high targeting fidelity.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序