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首页> 外文期刊>Biotechnology Journal: Healthcare,Nutrition,Technology >Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in-creased production capacity
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Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in-creased production capacity

机译：预选的CHO宿主细胞亚克隆的微阵列分析可鉴定与生产能力提高相关的基因表达模式

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Over the last three decades, product yields from CHO cells have increased dramatically, yet specific productivity (qP) remains a limiting factor. In a previous study, using repeated cell-sorting, we have established different host cell subclones that show superior transient qP over their respective parental cell lines (CHO-K1, CHO-S). The transcriptome of the resulting six cell lines in different biological states (untransfected, mock transfected, plasmid transfected) was first explored by hierarchical clustering and indicated that gene activity associated with increased qP did not stem from a certain cellular state but seemed to be inherent for a high qP host line. We then performed a novel gene regression analysis identifying drivers for an increase in qP. Genes significantly implicated were first systematically tested for enrichment of GO terms using a Bayesian approach incorporating the hierarchical structure of the GO term tree. Results indicated that specific cellular components such as nucleus, ER, and Golgi are relevant for cellular productivity. This was complemented by targeted GSA that tested functionally homogeneous, manually curated subsets of KEGG pathways known to be involved in transcription, translation, and protein processing. Significantly implicated pathways included mRNA surveillance, proteasome, protein processing in the ER and SNARE interactions in vesicular transport.
机译：在过去的三十年中，CHO细胞的产品产量急剧增加，但是比生产率（qP）仍然是一个限制因素。在先前的研究中，使用重复的细胞分类，我们建立了不同的宿主细胞亚克隆，它们的瞬时qP优于其各自的亲本细胞系（CHO-K1，CHO-S）。首先通过分层聚类探索了在不同生物学状态（未转染，模拟转染，质粒转染）中产生的六种细胞系的转录组，结果表明与增加的qP相关的基因活性并非源于某种细胞状态，而似乎是固有的高QP主机线。然后，我们进行了新颖的基因回归分析，确定了qP增加的驱动因素。首先，使用结合了GO术语树的层次结构的贝叶斯方法，系统地测试了显着牵连的基因对GO术语的富集。结果表明，特定的细胞成分（如核，ER和高尔基体）与细胞生产力有关。这由目标GSA补充，该目标GSA测试功能已知的KEGG途径的均质，人工策划的子集，该子集已知参与转录，翻译和蛋白质加工。显着牵连的途径包括mRNA监测，蛋白酶体，ER中的蛋白质加工和水泡运输中的SNARE相互作用。

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