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修改/脂质相关核酸 (PLANA) 用于CRISPR/Cas9核蛋白输送

Abdulelah Alhazza¹, Parvin Mahdipoor², Ryley Hall²

¹Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USA; Department of Pharmaceutics, Faculty of Pharmacy, Northern Border University, Rafha, 76313, Saudi Arabia.

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

|January 23, 2024

概括

化/脂质相关核酸 (PLANA) 是一种更安全,更有效的方法,可以将CRISPR/Cas9核核蛋白输送到细胞中进行基因组编辑. 这一进步解决了以前的毒性问题,为未来的体内应用铺平了道路.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
纳米技术在医学中的应用

背景情况:

使用集群定期间隔的短Palindromic重复 (CRISPR) 和CRISPR相关的蛋白9 (Cas9) 的基因组编辑使人们对用于蛋白沉默的核酸输送重新产生了兴趣.
将CRISPR/Cas9组件输入细胞核具有重大挑战,特别是与小干扰RNA (siRNA) 的细胞质输送相比.
之前的工作为siRNA传递引入了/脂相关核酸 (PLANAs),需要对CRISPR/Cas9应用进行进一步的研究.

研究的目的:

评估/脂相关核酸 (PLANA) 的效率和安全性,用于体外输送CRISPR/Cas9核糖蛋白.
通过引入聚乙烯糖醇 (PEG) 修改来解决与非PEGylated PLANAs观察到的毒性问题.

主要方法:

通过将PEG结合脂质纳入纳米粒子组合物中来实现PLANA的PEG化.
在PEGylated和非PEGylatedPLANAs中评估了体外传染效率和细胞内化.
量化了毒性,封装效率和表面PEGylation水平.
下一代测序用于分析转染细胞中的基因编辑结果 (indels).

主要成果:

PEGylation显著降低了PLANA的毒性,而不会显著损害封装效率.
PEGylated PLANAs的细胞内化和转染效率与非PEGylated版本和商业试剂 (LipofectamineTM CRISPRMAXTM) 相同.
下一代测序证实了转染细胞群体中各种内基的存在,表明成功的CRISPR/Cas9活动.

结论:

基化PLANA是一种有效和更安全的替代品,用于在体外输送CRISPR/Cas9核糖蛋白.
增强的PEGylated PLANA的安全性使得它们成为未来体内基因组编辑研究的有希望的候选者.
这项研究强调了纳米技术在增强基因编辑工具的交付方面的潜力.

关键词:

这就是CRISPR/Cas9的作用.在体外 (in vitro) 的情况下.脂质脂质脂质是一种一个计划,一个计划.聚乙烯糖醇是一种聚乙烯糖醇.转化转化转化转化

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