一个分裂和可诱导的腺基编辑器,用于精确的体内基编辑
Hongzhi Zeng1, Qichen Yuan1, Fei Peng2
1Department of Chemical and Biomolecular Engineering, Rice University, Houston, TX, 77005, USA.
Nature communications
|September 11, 2023
概括
研究人员开发了一种分裂腺基编辑器 (sABE),用于精确的DNA编辑. 这种新系统使用化学诱导的二分化来控制基编辑器活动,为潜在的治疗应用提供更高的精度和更少的非目标效应.
科学领域:
- 分子生物学分子生物学
- 基因编辑技术的技术
- 生物化学 生物化学
背景情况:
- 基因编辑器使得有针对性的核酸转换成为可能.
- 使用TadA脱氨酶的现有腺基编辑器 (ABE) 缺乏细胞控制.
- 对于精确的基因编辑应用,翻译后控制至关重要.
研究的目的:
- 开发一种具有化学诱导二元化 (CID) 的分裂腺基编辑器 (sABE),用于可控的DNA编辑.
- 与传统的ABE相比,评估sABE的活动,精度和非目标效应.
- 为了证明CID控制的基准编辑的in vivo适用性.
主要方法:
- 使用化学诱导二元化 (CID) 构建一个分裂腺基编辑器 (sABE) 系统.
- 在蜂模型中评估sABE的目标编辑效率和背景活动.
- 评估基因组和转录组非目标突变和编辑精度 (例如,单对双比率).
- 通过多重拼接供体破坏实现基因淘汰的SABE.
- 在小鼠模型中通过腺相关病毒 (AAV) 载体传递sABE进行体内编辑.
主要成果:
- sABE显示了高的目标编辑活动,与拉巴胺诱导后的ABE8e相美.
- 在没有诱导的情况下,sABE的背景活性显著降低.
- sABE通过更窄的活动窗口提高了精度,并改善了单对双腺素编辑比率.
- 通过sABE观察到减少了基因组和转录组的目标外影响.
- 在小鼠肝脏中的PCSK9基因中成功实现了基因淘汰和体内A•T转换为G•C基因.
结论:
- sABE为DNA基编辑活动提供了精确的,可诱导的控制.
- 增强的精度和减少的SABE的非目标效应有利于治疗应用.
- 使用sABE进行CID控制的基因编辑对基础研究和体内基因治疗都有很大的前景.
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