Engineering APOBEC3A Deaminase for Highly Accurate and Efficient Base Editing

Overview

Journal Nat Chem Biol

Specialties Biochemistry
Biology
Chemistry

Date 2024 Mar 30

PMID 38553609

Authors

Lei Yang

Yanan Huo

Man Wang

Dan Zhang

Tianai Zhang

Hao Wu

Xichen Rao

Haowei Meng

Shuming Yin

Jiale Mei

Dexin Zhang

Xi Chen

Jia Lv

Meizhen Liu

Yiyun Cheng

Yuting Guan

Bo Feng

Gaojie Song

Chengqi Yi

Mingyao Liu

Fanyi Zeng

Liren Wang

Dali Li

Affiliations

Soon will be listed here.

Abstract

Cytosine base editors (CBEs) are effective tools for introducing C-to-T base conversions, but their clinical applications are limited by off-target and bystander effects. Through structure-guided engineering of human APOBEC3A (A3A) deaminase, we developed highly accurate A3A-CBE (haA3A-CBE) variants that efficiently generate C-to-T conversion with a narrow editing window and near-background level of DNA and RNA off-target activity, irrespective of methylation status and sequence context. The engineered deaminase domains are compatible with PAM-relaxed SpCas9-NG variant, enabling accurate correction of pathogenic mutations in homopolymeric cytosine sites through flexible positioning of the single-guide RNAs. Dual adeno-associated virus delivery of one haA3A-CBE variant to a mouse model of tyrosinemia induced up to 58.1% editing in liver tissues with minimal bystander editing, which was further reduced through single dose of lipid nanoparticle-based messenger RNA delivery of haA3A-CBEs. These results highlight the tremendous promise of haA3A-CBEs for precise genome editing to treat human diseases.

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