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工程人类细胞表达CRISPR/Cas9-协同激活媒介用于重组蛋白质生产

Colby J Feser¹, James M Williams², Daniel T Lammers²

¹Department of Pediatrics, Division of Blood and Marrow Transplantation, MMC 366 Mayo, 8366A, 420 Delaware Street SE, Minneapolis, MN 55455, USA.

International journal of molecular sciences

|May 27, 2023

概括

将CRISPR/Cas9协同激活器调解器 (SAM) 系统集成到人体细胞中,可以实现强大的基因表达和蛋白质生产. 这种基因组集成的方法为重组工程和转录模拟提供了一个强大的工具.

科学领域:

分子生物学分子生物学
基因编辑基因编辑
生物技术是生物技术.

背景情况:

基于等离子体的蛋白质生产基因表达面临着翻译后修改和表达大型蛋白质的挑战.
克里斯普尔/卡斯9协同激活器调解器 (SAM) 系统提供可编程的转录模拟.
将SAM集成到人类基因组中,为基因表达提供了一个稳定的平台.

研究的目的:

研究将SAM系统集成到人类基因组中的有效性,以获得强大的基因表达和蛋白质生产.
为了证明基因组集成SAM对单重复和多重复基因向的能力.
突出SAM表达的人类细胞在重组工程和转录调制方面的潜力.

主要方法:

通过整合SAM系统组件来设计人类细胞系 (HEK293,HKB11,SK-HEP1,HEP-g2).
使用凝血因子X (FX) 和纤维素原 (FBN) 作为基因表达的概念验证标.
采用基于CRISPR/Cas9的SAM系统,包括dCas9与VP64,p65和HSF1转录激活器融合.

主要成果:

在多种人类细胞类型中实现了SAM系统的稳定集成.
在所有测试细胞系中观察到针对向基因的显著mRNA上调.
证明了同时发生的蛋白质表达,证实了成功的基因向和生产.

结论:

基因组集成的SAM系统为人类细胞中用户定义的基因表达和蛋白质生产提供了一个强大的平台.
这项技术使单复合和多复合基因向用于重组工程成为可能.
表达SAM的细胞具有广泛的基础研究,转化研究和临床应用的潜力.

关键词:

这就是CRISPR/Cas9的作用.细胞工程细胞工程凝血因子凝血因子纤维素原体是一种纤维原体.多重复杂的多重复杂.重组蛋白质蛋白质的重组蛋白质是什么?

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