Vito Davide, Eriksen Jens Christian, Skjødt Christian, Weilguny Dietmar, Rasmussen Søren K, Smales C Mark
Industrial Biotechnology Centre and School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK.
Symphogen A/S, Pederstrupvej 93, DK-2750 Ballerup, Denmark; AGC Biologics, Vandtårnsvej 83, DK-2860 Søborg, Denmark.
iScience. 2020 Jan 24;23(1):100785. doi: 10.1016/j.isci.2019.100785. Epub 2019 Dec 18.
How the long non-coding RNA (lncRNA) genome in recombinant protein producing Chinese hamster ovary (CHO) cell lines relates to phenotype is not well described. We therefore defined the CHO cell lncRNA transcriptome from cells grown in controlled miniature bioreactors under fed-batch conditions using RNA-Seq to identify lncRNAs and how the expression of these changes throughout growth and between IgG producers. We identify lncRNAs including Adapt15, linked to ER stress, GAS5, linked to mTOR signaling/growth arrest, and PVT1, linked to Myc expression, which are differentially regulated during fed-batch culture and whose expression correlates to productivity and growth. Changes in (non)-coding RNA expression between the seed train and the equivalent day of fed-batch culture are also reported and compared with existing datasets. Collectively, we present a comprehensive lncRNA CHO cell profiling and identify targets for engineering growth and productivity characteristics of CHO cells.
重组蛋白生产中国仓鼠卵巢（CHO）细胞系中的长链非编码RNA（lncRNA）基因组与细胞表型之间的关系尚未得到充分描述。因此，我们使用RNA测序技术，从在补料分批培养条件下的可控微型生物反应器中生长的细胞中定义了CHO细胞lncRNA转录组，以鉴定lncRNA，以及这些lncRNA在整个生长过程中以及在IgG生产细胞之间的表达变化情况。我们鉴定出了包括与内质网应激相关的Adapt15、与mTOR信号传导/生长停滞相关的GAS5以及与Myc表达相关的PVT1等lncRNA，它们在补料分批培养过程中受到差异调节，并且其表达与生产力和生长相关。我们还报告了种子批和补料分批培养相同天数之间（非）编码RNA表达的变化，并与现有数据集进行了比较。总体而言，我们展示了全面的CHO细胞lncRNA图谱，并确定了用于改造CHO细胞生长和生产力特性的靶点。
