Minimizing Clonal Variation during Mammalian Cell Line Engineering for Improved Systems Biology Data Generation,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Minimizing Clonal Variation during Mammalian Cell Line Engineering for Improved Systems Biology Data Generation
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-07-30 00:00:00 , DOI: 10.1021/acssynbio.8b00140
Lise Marie Grav ₁ , Daria Sergeeva ₁ , Jae Seong Lee _{1,

2} , Igor Marin de Mas ₁ , Nathan E. Lewis _{3,

4} , Mikael Rørdam Andersen ₅ , Lars Keld Nielsen _{1,

6} , Gyun Min Lee _{1,

7} , Helene Faustrup Kildegaard ₁

Affiliation

Mammalian cells are widely used to express genes for basic biology studies and biopharmaceuticals. Current methods for generation of engineered cell lines introduce high genomic and phenotypic diversity, which hamper studies of gene functions and discovery of novel cellular mechanisms. Here, we minimized clonal variation by integrating a landing pad for recombinase-mediated cassette exchange site-specifically into the genome of CHO cells using CRISPR and generated subclones expressing four different recombinant proteins. The subclones showed low clonal variation with high consistency in growth, transgene transcript levels and global transcriptional response to recombinant protein expression, enabling improved studies of the impact of transgenes on the host transcriptome. Little variation over time in subclone phenotypes and transcriptomes was observed when controlling environmental culture conditions. The platform enables robust comparative studies of genome engineered CHO cell lines and can be applied to other mammalian cells for diverse biological, biomedical and biotechnological applications.

中文翻译：

最小化哺乳动物细胞系工程设计过程中的克隆变异，以改善系统生物学数据的产生

哺乳动物细胞被广泛用于表达用于基础生物学研究和生物药物的基因。当前产生工程细胞系的方法引入了高基因组和表型多样性，这阻碍了基因功能的研究和新细胞机制的发现。在这里，我们通过使用CRISPR将整合重组酶介导的盒交换位点的着陆垫特异性整合到CHO细胞的基因组中，从而使克隆变异最小化，并产生了表达四种不同重组蛋白的亚克隆。亚克隆显示出低克隆变异性，在生长，转基因转录水平和对重组蛋白表达的整体转录反应方面具有高度一致性，从而能够更好地研究转基因对宿主转录组的影响。控制环境培养条件时，观察到亚克隆表型和转录组随时间的变化很小。该平台可对基因组改造的CHO细胞系进行可靠的比较研究，并可应用于其他哺乳动物细胞，用于多种生物学，生物医学和生物技术应用。

更新日期：2018-07-30

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11