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Myospreader通过核传播改善了骨肌肉中的基因编辑

Kiril K Poukalov^1,2,3, M Carmen Valero^1,2,3, Derek R Muscato^1,2,3

¹Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610.

Proceedings of the National Academy of Sciences of the United States of America

|April 30, 2024

概括

研究人员开发了"Myospreader",一种可以提高肌肉细胞CRISPR/Cas9基因编辑效率的. 这项创新增强了所有细胞核的蛋白质输送,提高了遗传肌肉疾病的治疗潜力.

科学领域:

分子生物学分子生物学
基因治疗基因治疗
肌肉生物学肌肉生物学

背景情况:

在骨肌中进行CRISPR/Cas9基因编辑需要有效地将Cas9输送到所有神经核中.
目前的核向基因疗法在超出初始的多核领域的货物传播方面面临限制.

研究的目的:

确定一种增强Cas9蛋白向肌纤维内的所有肌核传播的方法.
为了提高CRISPR/Cas9基因编辑在骨肌肉中的疗效,用于治疗应用.

主要方法:

核定位信号 (NLS) 和核出口信号 (NES) 的选,以确定有效的序列.
将已识别的序列,称为"Myospreader",附加到Cas9和dCas9蛋白质上.
在神经细胞和神经纤维中测试Myospreader修饰的Cas9,以测试蛋白质的稳定性和传播.
评估Myospreader-dCas9在肌性缩症小鼠模型中的转录抑制.
在CRISPR报告员和杜申肌肉衰竭小鼠模型中评估Myospreader-Cas9基因编辑效率.

主要成果:

鉴定出一种新组合的"myospreader"能够促进核细胞的传播.
Myospreader增强了Cas9蛋白的稳定性,并在体外和体内扩散到多个myonuclei.
在AAV交付的Myospreader dCas9中,在肌性衰变模型中,证明了对有毒RNA的改善抑制.
Myospreader-Cas9显著增加了杜氏肌肉发育不良和记者模型中的基因编辑率.

结论:

Myospreader代表了在增强核向基因疗法在骨肌肉中的交付方面的突破.
这种序列为改善核基因疗法中治疗载荷的空间分布提供了设计原则.
Myospreader强调了空间动态在开发有效的肌肉疾病基因疗法的关键作用.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?腺相关病毒的病毒.基因治疗的基因疗法多核领域的领域.空间基因调节基因调节

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