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开发可离子化脂质纳米颗粒和冷化配方,用于强大的CRISPR-Cas9传递和基因组编辑

Qian Sun¹, Hongqian Zhang¹, Feng Ding¹

¹Key Laboratory of Colloid and Interface Chemistry of the Ministry of Education, and School of Chemistry and Chemical Engineering, Shandong University, Jinan 25010, China.

International journal of pharmaceutics

|January 24, 2024

概括

研究人员开发了用于CRISPR-Cas9传递的新型电离性脂质纳米粒子 (iLNPs). 1A14-iLNP配方显示了基因编辑的高效率和mRNA疗法的潜力.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
基因编辑技术的技术

背景情况:

克里斯普尔-卡斯基因组编辑提供了巨大的生物医学潜力,但需要高效的传递系统.
开发有效的CRISPR-Cas组件,如Cas9mRNA和指导RNA (sgRNA) 的传递工具仍然是一个重大挑战.

研究的目的:

为了合成和评估新型的可电离性脂质纳米粒子 (iLNPs),用于CRISPR-Cas9mRNA和sgRNA的联合传递.
优化iLNP配方,以提高细胞吸收,内体逃生和基因编辑效率.
评估领先iLNP候选物的体内传递和生物相容性,并探索冷化以改善配方.

主要方法:

通过将烯酸与各种氨基分子结合,合成可离子化脂质.
电离性脂质纳米颗粒 (iLNPs) 的组装用于 Cas9 mRNA 和 sgRNA 的联合封装.
在体外评估细胞吸收,内体逃生和mRNA表达.
对基因编辑效率的评估,针对HeLa-EGFP细胞中的EGFP.
针对肝脏向输送和生物相容性评估的体内研究.
开发一种由冷化介导的逆转染法.

主要成果:

1A14-iLNP配方,利用脂质与精子作为氨基头,在体外表现出优异的细胞吸收,内体逃生和mRNA表达.
使用1A14-iLNP同时输送Cas9mRNA和sgRNA,在HeLa-EGFP细胞中达到高达70%的EGFP淘汰效率.
在体内研究表明,1A14-iLNP通过良好的生物相容性向肝脏准Cas9mRNA的被动输送.
为了高效的CRISPR-Cas9传递,建立了一个简单的冷化中介反转染法.

结论:

新的1A14-iLNP配方代表了CRISPR-Cas9传递的高效系统,改善了基因编辑结果.
开发的iLNP和冷化技术为推进基于mRNA和CRISPR的治疗应用提供了一个有希望的解决方案.
这项工作通过增强的交付策略,促进了CRISPR-Cas技术的更广泛的生物医学应用.

关键词:

克里斯普尔-Cas9是什么意思基因组编辑基因组编辑脂质纳米颗粒是一种纳米粒子.软化液化化是什么意思提供mRNA的交付.

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