双链断裂修复途径通过双AAV向量对处理和转导产生不同的影响
Anna C Maurer1,2, Brian Benyamini1, Vinson B Fan1
1Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
bioRxiv : the preprint server for biology
|October 4, 2023
概括
抑制DNA修复途径,如同源复合 (HR),通过使用双重复合腺相关病毒载体 (rAAV) 增强基因传递. 这改善了大型转基因的表达,克服了rAAV的大小限制.
科学领域:
- 分子生物学分子生物学
- 基因治疗 基因治疗
- 病毒学 病毒学
背景情况:
- 复合腺相关病毒载体 (rAAV) 广泛用于基因传递.
- 目前的rAAV载体具有有限的DNA承载能力,限制了治疗应用.
- 双转接向量通过连接rAAV基因组来扩大有效载荷,但效率是可变的.
研究的目的:
- 为了确定影响双 rAAV 矢量传导效率的宿主因素.
- 研究宿主细胞调节器在rAAV基因组连接中的作用.
- 探索使用rAAV.rAAV增强大基因有效载荷传递的策略.
主要方法:
- 全基因组屏幕用于识别双向量转导的宿主细胞调节器.
- 减少和抑制关键宿主因子,包括BRCA1和Rad51.
- 在rAAV分娩后评估转基因复制和表达.
主要成果:
- 同源重组 (HR) 途径被确定为抑制双重rAAV转导.
- 减少或抑制HR因子BRCA1和Rad51显著增加了转基因复制.
- 在HR抑制后观察到增强的连接和大分裂转基因的表达.
结论:
- 主体DNA损伤修复途径,特别是HR,在rAAV转导效率中起着至关重要的作用.
- 针对HR因素提供了一种新的策略,以提高rAAV载体的基因有效载荷能力.
- 药理上抑制HR是一种增强基因疗法的有希望的方法.
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