Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, Penang, Malaysia.
Mol Biotechnol. 2020 Sep;62(9):401-411. doi: 10.1007/s12033-020-00265-9. Epub 2020 Aug 4.
A broad range of cell lines with characteristic features are used as bio-factories to produce recombinant proteins for basic research and therapeutic purposes. Genetic engineering strategies have been used to manipulate the genome of mammalian cells, insects, and yeasts for heterologous expression. One reason is that the glycosylation pattern of the expression hosts differs somehow from mammalian cells, which may cause immunogenic reactions upon administration in humans. CRISPR-Cas9 is a simple, efficient, and versatile genome engineering tool that can be programmed to precisely make double-stranded breaks at the desired loci. Compared to the classical genome editing methods, a CRISPR-Cas9 system is an ideal tool, providing the opportunity to integrate or delete genes from the target organisms. Besides broadened applications, limited studies have used CRISPR-Cas9 for editing the endogenous pathways in expression systems for biopharmaceutical applications. In the present review, we discuss the use of CRISPR-Cas9 in expression systems to improve host cell lines, increase product yield, and humanize glycosylation pathways by targeting intrinsic genes.
广泛的具有特征的细胞系被用作生物工厂,以生产用于基础研究和治疗目的的重组蛋白。遗传工程策略已被用于操纵哺乳动物细胞、昆虫和酵母的基因组,以进行异源表达。原因之一是表达宿主的糖基化模式在某种程度上与哺乳动物细胞不同,这可能会导致在人类中给药时产生免疫原性反应。CRISPR-Cas9 是一种简单、高效、多功能的基因组工程工具,可以编程在所需的基因座上精确地产生双链断裂。与经典的基因组编辑方法相比,CRISPR-Cas9 系统是一种理想的工具,为从目标生物中整合或删除基因提供了机会。除了应用范围的扩大,有限的研究已经使用 CRISPR-Cas9 编辑表达系统中的内源性途径,用于生物制药应用。在本综述中,我们讨论了使用 CRISPR-Cas9 来改善宿主细胞系、提高产物产量以及通过靶向内在基因使糖基化途径人源化,从而提高生物制药产品的质量。