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有效的基因组编辑使用"纳米医学"AsCas12a-VLPs生产与Pol II转录的crRNA

Sofiia E Borovikova¹, Mikhail V Shepelev², Dmitriy V Mazurov²

¹Institute of Gene Biology Russian Academy of Sciences, 119334 Moscow, Russia.

International journal of molecular sciences

|December 17, 2024

概括

这项研究开发了使用AsCas12a.a进行基因编辑的先进病毒样粒子 (VLP). 这些VLP有效地将基因编辑工具传送到细胞中,提高治疗应用的淘汰效率.

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:

病毒样粒子 (VLPs) 被探索为提供基因编辑组件,如Cas核酶和指导RNA核糖核蛋白复合体 (RNP).
目前使用SpCas9和U6促销器驱动的sgRNA的方法面临着由于sgRNA的核定位而面临的挑战,阻碍了有效的RNP包装到细胞质VLP中.

研究的目的:

设计VLP以有效地传递AsCas12a,并引导RNA (crRNA) 进行增强的基因编辑.
通过优化RNP复杂形成和包装来克服以前的VLP系统的局限性.

主要方法:

使用"NanoMEDIC"VLP系统包装AsCas12a及其相关crRNA.
采用RNA聚合酶II (Pol II) 驱动的CMV促进剂用于crRNA的细胞质表达,从而使AsCas12a.a.能够高效地处理.
对哺乳动物细胞的基因编辑效率测试单个和排列crRNAs.

主要成果:

与SpCas9-VLP相比,AsCas12a-VLP显著改善了RNP整合和基因编辑效率.
在293个细胞中,CMV驱动的crRNA增加了转基因淘汰 (金星,CCR5) 从30%增加到50-90%.
在Jurkat T细胞中,内源性CXCR4淘汰从单个crRNA增加了1%至20%,在crRNA数组中增加到60-70%.

结论:

AsCas12a和CMV驱动的crRNA可以有效地被包装到VLPs中,通过中介高级基因编辑.
AsCas12a-VLPs代表了一种有希望的新工具,用于在哺乳动物细胞中提供RNP,用于基因编辑和潜在的基因治疗应用.

关键词:

这就是AsCas12a.在CRISPR/Cas交付过程中.纳米医学 (NanoMEDIC) 是一个这是Pol II的推广者.核蛋白复合体的组合物.类似病毒的颗粒.

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相关概念视频

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...