通过CRISPR引导的DNA基编辑器诱导的转录组范围内的非目标RNA编辑
Julian Grünewald1,2,3,4, Ronghao Zhou1,2,3, Sara P Garcia1
1Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA.
Nature
|April 18, 2019
概括
细胞因子基编辑器 (CBEs) 可以无意中改变RNA细胞因子,导致人类细胞中广泛的转录组编辑. 设计的CBE变体显著减少了这些非目标RNA编辑,同时保持了DNA编辑精度.
科学领域:
- 分子生物学
- 遗传学
- 生物技术
背景情况:
- CRISPR-Cas基编辑使得针对研究和治疗的DNA变化成为可能.
- 细胞基编辑器 (CBEs) 使用APOBEC1进行C-T转换.
- 之前的研究发现CBE的DNA编辑异常.
研究的目的:
- 研究CBE在人体细胞中的潜在非目标RNA编辑.
- 设计具有减少RNA编辑活动的CBE变体.
- 评估RNA编辑对基因功能和蛋白质产品的影响.
主要方法:
- 在人类细胞中使用基于老鼠APOBEC1的CBE系统.
- 进行全转录组分析以识别C-U编辑.
- 为减少RNA编辑工程和测试的突变APOBEC1变体.
- 评估目标DNA编辑效率和工程变体的特异性.
主要成果:
- 在人类细胞中诱导了广泛的转录组C-to-U编辑,影响了很大比例的基因.
- 在编码和非编码区域发生了RNA编辑,导致各种突变.
- 工程化CBE变种显示RNA编辑减少了390倍和3800倍.
- 突变的CBE在没有显著的效率损失的情况下表现出更好的目标DNA编辑精度.
- 还发现腺基编辑器可诱导全转录组RNA编辑.
结论:
- CBE可能会导致无意的,广泛的RNA修饰.
- 工程工作可以产生具有增强特异性的基编辑器,并减少RNA的目标外影响.
- 对于基编辑器在研究和临床中的应用而言,对RNA脱效应的彻底描述至关重要.
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