Decreasing predictable DNA off-target effects and narrowing editing windows of adenine base editors by fusing human Rad18 protein variant

被引：1

作者：

Wang, Ziru ^{[1
]}

Yuan, Hongming ^{[1
,2
,3
]}

Yang, Lin ^{[1
]}

Ma, Lerong ^{[1
]}

Zhang, Yuanzhu ^{[1
]}

Deng, Jiacheng ^{[1
]}

Li, Xueyuan ^{[1
]}

Xiao, Wenyu ^{[1
]}

Li, Zhanjun ^{[1
]}

Qiu, Jiazhang ^{[4
]}

Ouyang, Hongsheng ^{[1
,2
,3
]}

Pang, Daxin ^{[1
,2
,3
]}

机构：

[1] Jilin Univ, Coll Anim Sci, Changchun 130062, Peoples R China

[2] Jilin Univ, Chongqing Res Inst, Chongqing 401123, Peoples R China

[3] Chongqing Jitang Biotechnol Res Inst, Chongqing 401123, Peoples R China

[4] Jilin Univ, Coll Vet Med, Changchun 130062, Peoples R China

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2023年 / 253卷

基金：

中国国家自然科学基金;

关键词：

Base editors; RING type E3 ubiquitin ligase; Editing windows; Off-target; GENOMIC DNA; CRISPR; REPAIR; STRAND; PCSK9;

D O I：

10.1016/j.ijbiomac.2023.127418

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Adenine base editors, enabling targeted A-to-G conversion in genomic DNA, have enormous potential in ther-apeutic applications. However, the currently used adenine base editors are limited by wide editing windows and off-target effects in genetic therapy. Here, we report human e18 protein, a RING type E3 ubiquitin ligase variant, fusing with adenine base editors can significantly improve the preciseness and narrow the editing windows compared with ABEmax and ABE8e by diminishing the abundance of base editor protein. As a proof of concept, ABEmax-e18 and ABE8e-e18 dramatically decrease Cas9-dependent and Cas9-independent off-target effects than traditional adenine base editors. Moreover, we utilized ABEmax-e18 to establish syndactyly mouse models and achieve accurate base conversion at human PCSK9 locus in HepG2 cells which exhibited its potential in genetic therapy. Furthermore, a truncated version of base editors-RING (ABEmax-RING or AncBE4max-RING), which fusing the 63 amino acids of e18 protein RING domain to the C terminal of ABEmax or AncBE4max, exhibited similar effect compared to ABEmax-e18 or AncBE4max-e18.In summary, the e18 or RING protein fused with base editors strengthens the precise toolbox in gene modification and maybe works well with various base editing tools with a more applicable to precise genetic therapies in the future.

引用

页数：12