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包装的CRISPR-Cas9核糖蛋白加速了基因组编辑

Hannah Karp^1,2, Madeline Zoltek³, Kevin Wasko^2,3

¹Department of Chemistry, University of California, Berkeley, CA 94720, United States.

Nucleic acids research

|March 4, 2025

概括

通过包裹式输送车辆 (EDV) 输送CRISPR-Cas9核核蛋白 (RNP) 的效率是电穿孔的30倍以上. 这种包装的输送方法显著降低了有效的基因组编辑所需的剂量.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
生物技术是生物技术.

背景情况:

有效的CRISPR-Cas9基因组编辑需要将核糖蛋白 (RNP) 最佳地输送到细胞中.
目前的临床应用主要依赖于用于RNP输送的电穿孔,对替代方法的比较数据有限.
尽量减少免疫反应,非目标效应和细胞毒性对于安全有效的基因组编辑至关重要.

研究的目的:

系统地比较用于CRISPR-Cas9 RNP传递的电穿孔和封装输送车辆 (EDV).
用不同的传递策略来确定基因组编辑中Cas9 RNPs的剂量要求.
调查交付方法对编辑效率,动力学和RNP核停留时间的影响.

主要方法:

两个CRISPR-Cas9 RNP传递策略的比较:电穿孔和EDV.
使用光相关谱法量化Cas9 RNP剂量要求.
评估每个交付方法的基因组编辑效率和动力学.

主要成果:

有效的基因组编辑每核需要超过1300个Cas9 RNP.
通过EDV介导的基因组编辑比电穿孔效率高30倍以上.
在类似的RNP剂量下,EDV传递的编辑动态与电穿孔相比,至少是编辑动态的2倍.

结论:

像EDV这样的包装配送车辆为CRISPR-Cas9 RNP配送提供了比电穿孔更有效的替代方案.
通过长时间的RNP核居住,EDV交付可以提高编辑结果.
这项研究突出了EDVs在减少CRISPR-Cas9 RNP剂量所需的实验和临床基因组编辑应用中的潜力.

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