细胞因子基编辑器的工程与DNA损伤最小化和编辑范围多样化编辑范围多样化
Bo Yuan1,2, Shuqian Zhang1,3, Liting Song4,5
1Institute of Pediatrics, National Children's Medical Center, Children's Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200032, China.
Nucleic acids research
|October 16, 2023
概括
细胞因子基编辑器 (CBEs) 可以导致DNA双链断裂 (DSB),这是一个安全风险. 这项研究设计了更安全的CBE变体,具有降低的基因毒性,并扩展了基因编辑应用程序的编辑功能.
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:
- 细胞因子基编辑器 (CBEs) 提供精确的C-to-T替代,但面临安全问题.
- 非目标DNA/RNA编辑和基因毒性,特别是DNA双链断裂 (DSB),限制了当前的CBE应用.
- 现有的策略已经提高了DNA/RNA的准确性,但DSB风险评估和缓解仍然是关键的挑战.
研究的目的:
- 评估和最小化与DSB相关的CBE的DNA损伤风险.
- 设计新的CBE变体,以增强安全配置和扩大编辑范围.
- 开发一个更新的平台来评估CBE诱导的基因毒性.
主要方法:
- 设计了一种CBE变体 (YE1),并通过γH2AX积累评估其DNA/RNA的目标外编辑和DSB相关的DNA损伤.
- 在鱼LjCDA1和人类APOBEC3A上进行了除 aminase工程,以创建新的CBEs.
- 利用内部融合策略来多样化APOBEC3A衍生CBE的编辑范围和序列偏好.
主要成果:
- YE1 CBE变种诱导了人类细胞中与DSB相关的显著DNA损伤.
- 工程LjCDA1和APOBEC3A衍生的CBEs证明了消除了DSB风险,并最大限度地减少了目标之外的编辑.
- 来自APOBEC3A的CBE通过内部融合显示了多样化的编辑范围和序列偏好.
结论:
- 本研究为CBE诱导的DSB风险提供了一个改进的评估平台.
- 产生了新的CBE工具包,其基因毒性降低,编辑签名多样化.
- 这些进展旨在扩大CBE在基因编辑中的安全应用.
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