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基于共感染和CRISPR/Cas9的昆虫细胞中诱导性淘汰系统的评估

Christina Sophie Hausjell¹, Miriam Klausberger¹, Wolfgang Ernst¹

¹Department of Biotechnology, Institute of Molecular Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.

|July 27, 2023

概括

这项研究引入了CRISPR/Cas9系统,用于减少VLP生产中不必要的细菌病毒. 针对重要基因显著降低病毒标位,帮助生物制药净化过程.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
病毒学病毒学

背景情况:

细菌病毒/昆虫细胞系统是一个具有成本效益的平台,用于生产复杂的蛋白质,如病毒样颗粒 (VLPs).
在VLP生产过程中芽的baculovirus共同形成使下游加工复杂化,并增加成本.
VLP是对新兴病毒威胁的有希望的疫苗候选者.

研究的目的:

开发一种可诱导的淘汰系统,以减少VLP生产中的芽百科病毒.
评估针对关键的细菌病毒基因 (gp64和vp80) 进行淘汰的有效性.

主要方法:

利用CRISPR/Cas9技术与两个共感染的细菌病毒载体进行诱导性基因淘汰.
设计的载体可以分别输送Cas9核酶和sgRNA.
向的重要细菌病毒基因 (gp64,vp80) 参与了芽.

主要成果:

通过向gp64和vp80.0,通过向gp64和vp80.0实现了百分之九十以上的细菌病毒标位降低.
由于基因淘汰,观察到重组蛋白质生产 (eYFP光强度) 的下降.
证明成功诱导基本的baculoviral基因的淘汰.

结论:

开发的CRISPR/Cas9系统有效地降低了VLP生产中的芽百科病毒标位.
工程和净化策略的进一步优化是必要的,以平衡病毒减少与重组蛋白质产量.
这种方法有可能提高基于VLP的疫苗生产的成本效益和效率.

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