Objectives

We used miRNA and proteomic profiling to understand intracellular pathways that contribute to high and low specific productivity (Qp) phenotypes in CHO clonally derived cell lines (CDCLs) from the same cell line generation project.

Results

Differentially expressed (DE) miRNAs were identified which are predicted to target several proteins associated with protein folding. MiR-200a was found to have a number of predicted targets associated with the unfolded protein response (UPR) which were shown to have decreased expression in high Qp CDCLs and have no detected change at the mRNA level. MiR-200a overexpression in a CHO CDCL was found to increase recombinant protein titer by 1.2 fold and Qp by 1.8 fold.

Conclusion

These results may suggest a role for miR-200a in post-transcriptional regulation of the UPR, presenting miR-200a as a potential target for engineering industrially attractive CHO cell phenotypes.

中文翻译：

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miRNA 的差异表达和 mir-200a 在表达 Fc 融合蛋白的高产和低产 CHO 细胞中的功能作用

目标

我们使用 miRNA 和蛋白质组学分析来了解导致来自同一细胞系生成项目的 CHO 克隆衍生细胞系 (CDCL) 中高和低比生产率 (Qp) 表型的细胞内途径。

结果

鉴别出差异表达 (DE) miRNA，预测其靶向与蛋白质折叠相关的几种蛋白质。发现 MiR-200a 具有许多与未折叠蛋白反应 (UPR) 相关的预测靶标，这些靶标显示在高 Qp CDCL 中表达降低，并且在 mRNA 水平上没有检测到变化。发现在 CHO CDCL 中过表达 MiR-200a 使重组蛋白滴度增加 1.2 倍，Qp 增加 1.8 倍。

结论

这些结果可能表明 miR-200a 在 UPR 的转录后调节中发挥作用，将 miR-200a 作为工程化具有工业吸引力的 CHO 细胞表型的潜在靶标。