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对于rAAV的产生,CRISPR屏幕涉及与感染相关的基因

Emily E O'Driscoll^1,2,3, Sakshi Arora^1,4,3, Jonathan F Lang^1,4

¹Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.

bioRxiv : the preprint server for biology

|September 30, 2024

概括

基因疗法制造是昂贵的. 研究人员使用CRISPR查,在生产细胞中发现基因,当它们被淘汰时,会增加复合腺相关病毒 (rAAV) 的产量,从而可能降低基因治疗成本.

科学领域:

分子生物学分子生物学
基因治疗制造业基因治疗制造业
病毒学病毒学

背景情况:

重组腺相关病毒 (rAAV) 载体对于基因疗法至关重要,有几种已批准的治疗方法,还有更多在开发中.
目前的rAAV制造过程昂贵,耗时,劳动密集,阻碍了更广泛的临床应用.
识别可以增强rAAV生产的宿主因素对于提高制造效率至关重要.

研究的目的:

确定可以向增加重组腺相关病毒 (rAAV) 颗粒产量的宿主基因.
探索宿主细胞机械在病毒载体生产和组装中的作用.
制定更高效和更具成本效益的rAAV制造战略.

主要方法:

在HEK 293细胞中进行了全基因组的CRISPR淘汰屏幕,以确定影响rAAV生产的基因.
采用了针对完整的AAV2囊体的抗体和高通量查的流细胞计.
分析了特定基因淘汰对rAAV产量的影响.

主要成果:

敲除以前与rAAV感染性相关的肝硫酸乙生物合成基因,降低了rAAV的产生.
几种囊泡贩运蛋白质的破坏显著增加了生产细胞中的rAAV产量.
证明了参与病毒感染的宿主蛋白可以被重新用于病毒组装.

结论:

宿主细胞的基因操纵可以增强复合腺相关病毒 (rAAV) 的产生.
膀性贩运途径代表了优化病毒载体制造的潜在目标.
这些发现有助于开发更高效,更可扩展的基因疗法生产方法.

关键词:

在 AAV AAV AAV 中.在CRISPR屏幕上使用CRISPR.基因治疗的基因疗法 rAAV2 rAAV2 的时间病毒的生产病毒的生产.

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