在胞嘧啶碱基编辑器中，UGI在Cas9内的重新定位减少了Cas9依赖的脱靶效应。

IF 3.9 2区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Scientific Reports Pub Date : 2025-10-10 DOI:10.1038/s41598-025-19482-w

Zehao Shi, Tian-Lin Cheng

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引用次数: 0

摘要

胞嘧啶碱基编辑器（CBEs）通过添加尿嘧啶DNA糖基酶抑制剂（UGI）与Cas9镍酶（nCas9）和胞苷脱氨酶实现精确的c -t转换，在nCas9羧基端进行常规融合，有效抑制尿嘧啶切除修复，提高编辑效率。然而，尽管具有强大的靶标活性，经典cbe表现出显著的cas9依赖性DNA脱靶效应，这需要对未来的应用进行优化。在这里，我们通过nCas9架构的内部融合提出了一个战略性的UGI迁移。这种空间重组保持了相当的靶上编辑效率，同时大大降低了cas9依赖性DNA的脱靶活性。我们的研究结果为开发高保真cbe建立了另一种工程范例，为广泛的基因组编辑应用提供了改进的平台。

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UGI relocation inside Cas9 reduces Cas9 dependent off target effects in cytosine base editors.

Cytosine base editors (CBEs) achieve precise C-to-T conversions by addition of uracil DNA glycosylase inhibitor (UGI) with Cas9 nickase (nCas9) and cytidine deaminase, and the conventional fusion at the nCas9 carboxyl terminus effectively inhibits uracil excision repair to enhance editing efficiency. However, despite potent on-target activity, classical CBEs exhibit significant Cas9-dependent DNA off-target effects that necessitate optimization for future applications. Here we present a strategic UGI relocation through internal fusion within the nCas9 architecture. This spatial reorganization maintains comparable on-target editing efficiency while substantially reducing Cas9-dependent DNA off-target activity. Our findings establish an alternative engineering paradigm to develop high-fidelity CBEs, offering an improved platform for widespread genome editing applications.

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来源期刊

Scientific Reports Natural Science Disciplines-

CiteScore

7.50

自引率

4.30%

发文量

19567

审稿时长

3.9 months

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