通过脂质介导的CRISPR/Cas9核糖蛋白复合物的输送用于基因编辑和纠正
Mert Öktem1, Thai Hoang Nguyen1, Esmeralda D C Bosman1
1Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Faculty of Science, Utrecht University, Utrecht 3584 CG, the Netherlands.
概括
研究人员开发了用于CRISPR/Cas9基因编辑传递的新型脂肪酸修饰载体. 用油酸修饰的LAH5纳米复合体在小鼠中表现出卓越的性能和体内基因编辑,显示出对遗传疾病的治疗潜力.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 基因工程是一种基因工程.
背景情况:
- CRISPR/Cas基因编辑为遗传疾病提供了潜在的治疗方法.
- 对CRISPR/Cas9核糖核蛋白 (RNP) 复合物的安全有效的细胞内传递仍然是一个挑战.
- 之前的研究使用了细胞透 (CPP) LAH5与Cas9 RNP和DNA修复模板进行交付.
研究的目的:
- 测试和描述LAH5-Cas9 RNP载体的新型脂肪酸修饰版本.
- 为了评估脂肪酸修改对RNP/纳米复合体性能的影响.
- 评估优化输送系统的体内疗效和安全性.
主要方法:
- 合成和表征了各种脂肪酸修饰的LAH5-Cas9 RNP纳米复合物.
- 评估了纳米复合体在血清中的稳定性,膜破坏和体外传染疗效.
- 在体外评估基因编辑和校正效率.
- 确定了对蛋白质酶消化的保护作用.
- 在转基因小鼠体内进行了体内基因编辑研究,通过肌肉内注射.
主要成果:
- 脂肪酸修饰改善了纳米复合体的稳定性,膜破坏和传染功效.
- Cas9 RNP/油酸LAH5纳米复合体表现出最佳的基因编辑和纠正.
- 这些纳米复合体保护了Cas9蛋白免受蛋白酶降解.
- 在体内研究中,在骨肌中实现了约10%的基因编辑,准CAG-tdTomato,约7%的准Ccr5,没有可观察到的短期毒性.
结论:
- 油酸修饰的LAH5作为直接Cas9/RNP输送的有效输送平台.
- 这种优化交付系统显示了基于CRISPR/Cas9的治疗应用的巨大潜力.
- 进一步的开发可以促进各种遗传疾病的治疗.
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