Specific Activation of the Expression of Growth Factor Genes in Expi293F Human Cells Using CRISPR/Cas9-SAM Technology Increases Their Proliferation.

IF 2 4区 生物学 Q4 CELL BIOLOGY
P A Bobrovsky, E N Grafskaia, D D Kharlampieva, V A Manuvera, V N Lazarev
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引用次数: 0

Abstract

Human cell lines play an important role in biotechnology and pharmacology. For them to grow, they need complex nutrient media containing signaling proteins - growth factors. We have tested a new approach that reduces the need of cultured human cell lines for exogenous growth factors. This approach is based on the generation of a modified cell with a selectively activated gene expression of one of the endogenous growth factors: IGF-1, FGF-2, or EIF3I. We modified the Expi293F cell line, a HEK293 cell line variant widely used in the production of recombinant proteins. Gene expression of the selected growth factors in these cells was activated using CRISPR/Cas9 technology with the synergistic activation mediators CRISPR/Cas9-SAM, which increased the expression of the selected genes at both mRNA and protein levels. Upon culturing under standard conditions, the modified lines exhibited increased proliferation. A synergistic effect was observed in co-culture of the three modified lines. In our opinion, these results indicate that this approach is promising for efficient modification of cell lines used in biotechnology.

利用 CRISPR/Cas9-SAM 技术特异性激活 Expi293F 人体细胞中生长因子基因的表达可增加其增殖。
人类细胞系在生物技术和药理学中发挥着重要作用。它们需要含有信号蛋白(生长因子)的复杂营养介质才能生长。我们对一种新方法进行了测试,这种方法可以减少培养人体细胞系对外源性生长因子的需求。这种方法的基础是产生一种改造细胞,其基因表达选择性地激活一种内源性生长因子:IGF-1、FGF-2 或 EIF3I。我们改造了 Expi293F 细胞系,这是一种广泛用于生产重组蛋白的 HEK293 细胞系变体。利用 CRISPR/Cas9 技术和协同激活介质 CRISPR/Cas9-SAM,激活了这些细胞中选定生长因子的基因表达,从而提高了选定基因在 mRNA 和蛋白质水平上的表达。在标准条件下培养时,改造品系的增殖速度加快。在三种改造品系的联合培养中观察到了协同效应。我们认为,这些结果表明,这种方法有望有效改造生物技术中使用的细胞系。
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来源期刊
Acta Naturae
Acta Naturae 农林科学-林学
CiteScore
3.50
自引率
5.00%
发文量
0
审稿时长
>12 weeks
期刊介绍: Acta Naturae is an international journal on life sciences based in Moscow, Russia. Our goal is to present scientific work and discovery in molecular biology, biochemistry, biomedical disciplines and biotechnology. These fields represent the most important priorities for the research and engineering development both in Russia and worldwide. Acta Naturae is also a periodical for those who are curious in various aspects of biotechnological business, innovations in pharmaceutical areas, intellectual property protection and social consequences of scientific progress. The journal publishes analytical industrial surveys focused on the development of different spheres of modern life science and technology. Being a radically new and totally unique journal in Russia, Acta Naturae is useful to both representatives of fundamental research and experts in applied sciences.
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