Domain-inlaid Nme2Cas9 adenine base editors with improved activity and targeting scope

被引：5

作者：

Bamidele, Nathan ^{[1
]}

Zhang, Han ^{[1
]}

Dong, Xiaolong ^{[2
]}

Cheng, Haoyang ^{[1
]}

Gaston, Nicholas ^{[1
]}

Feinzig, Hailey ^{[1
]}

Cao, Hanbing ^{[1
]}

Kelly, Karen ^{[1
]}

Watts, Jonathan K. ^{[1
,3
,4
]}

Xie, Jun ^{[5
,6
,7
,8
]}

Gao, Guangping ^{[5
,6
,7
,8
]}

Sontheimer, Erik J. ^{[1
,8
,9
]}

机构：

[1] Univ Massachusetts, RNA Therapeut Inst, Chan Med Sch, Worcester, MA 01605 USA

[2] Tessera Therapeut, Somerville, MA 02143 USA

[3] Univ Massachusetts, Dept Biochem & Mol Biotechnol, Chan Med Sch, Worcester, MA 01605 USA

[4] Univ Massachusetts, NeuroNexus Inst, Chan Med Sch, Worcester, MA 01605 USA

[5] Univ Massachusetts, Horae Gene Therapy Ctr, Chan Med Sch, Worcester, MA 01605 USA

[6] Univ Massachusetts, Chan Med Sch, Viral Vector Core, Worcester, MA 01605 USA

[7] Univ Massachusetts, Dept Microbiol & Physiol Syst, Chan Med Sch, Worcester, MA 01605 USA

[8] Univ Massachusetts, Li Weibo Inst Rare Dis Res, Chan Med Sch, Worcester, MA 01605 USA

[9] Univ Massachusetts, Program Mol Med, Chan Med Sch, Massachusetts, MA 01605 USA

来源：

NATURE COMMUNICATIONS | 2024年 / 15卷 / 01期

基金：

美国国家卫生研究院;

关键词：

GENOMIC DNA; CRISPR-CAS9; CAS9;

D O I：

10.1038/s41467-024-45763-5

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Nme2Cas9 has been established as a genome editing platform with compact size, high accuracy, and broad targeting range, including single-AAV-deliverable adenine base editors. Here, we engineer Nme2Cas9 to further increase the activity and targeting scope of compact Nme2Cas9 base editors. We first use domain insertion to position the deaminase domain nearer the displaced DNA strand in the target-bound complex. These domain-inlaid Nme2Cas9 variants exhibit shifted editing windows and increased activity in comparison to the N-terminally fused Nme2-ABE. We next expand the editing scope by swapping the Nme2Cas9 PAM-interacting domain with that of SmuCas9, which we had previously defined as recognizing a single-cytidine PAM. We then use these enhancements to introduce therapeutically relevant edits in a variety of cell types. Finally, we validate domain-inlaid Nme2-ABEs for single-AAV delivery in vivo. Nme2Cas9 has been well established as a genome editing platform. Here the authors engineer Nme2Cas9 to further increase the activity and targeting scope of compact Nme2Cas9 base editors and validate domain-inlaid Nme2-ABEs for single-AAV delivery in vivo.

引用

页数：13

共 48 条

[1] Genome editing with CRISPR-Cas nucleases, base editors, transposases and prime editors [J].