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

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dc.citation.articleNumbereaba1773
dc.citation.issueNumber29
dc.citation.journalTitleScience Advances
dc.citation.volumeNumber6
dc.contributor.authorLee, Sangsin
dc.contributor.authorDing, Ning
dc.contributor.authorSun, Yidi
dc.contributor.authorYuan, Tanglong
dc.contributor.authorLi, Jing
dc.contributor.authorYuan, Qichen
dc.contributor.authorLiu, Lizhong
dc.contributor.authorYang, Jie
dc.contributor.authorWang, Qian
dc.contributor.authorKolomeisky, Anatoly B.
dc.contributor.authorHilton, Isaac B.
dc.contributor.authorZuo, Erwei
dc.contributor.authorGao, Xue
dc.contributor.orgCenter for Theoretical and Biological Physics
dc.date.accessioned2020-10-30T19:43:45Z
dc.date.available2020-10-30T19:43:45Z
dc.date.issued2020
dc.description.abstractCytosine 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.
dc.identifier.citationLee, Sangsin, Ding, Ning, Sun, Yidi, et al.. "Single C-to-T substitution using engineered APOBEC3G-nCas9 base editors with minimum genome- and transcriptome-wide off-target effects." <i>Science Advances,</i> 6, no. 29 (2020) American Association for the Advancement of Science: https://doi.org/10.1126/sciadv.aba1773.
dc.identifier.doihttps://doi.org/10.1126/sciadv.aba1773
dc.identifier.urihttps://hdl.handle.net/1911/109465
dc.language.isoeng
dc.publisherAmerican Association for the Advancement of Science
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/
dc.titleSingle C-to-T substitution using engineered APOBEC3G-nCas9 base editors with minimum genome- and transcriptome-wide off-target effects
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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