差异表达的 miRNAs 和 mir-200a 在高和低表达 Fc 融合蛋白的 CHO 细胞中的功能作用。
Differential expression of miRNAs and functional role of mir-200a in high and low productivity CHO cells expressing an Fc fusion protein.
机构信息
National Institute for Cellular Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.
National Institute for Bioprocessing Research and Training, Dublin 4, Ireland.
出版信息
Biotechnol Lett. 2021 Aug;43(8):1551-1563. doi: 10.1007/s10529-021-03153-7. Epub 2021 Jun 16.
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.
目的
我们使用 miRNA 和蛋白质组学分析来了解细胞内途径，这些途径有助于 CHO 克隆衍生细胞系 (CDCL) 中高和低比生产率 (Qp) 表型的形成，这些 CDCL 来自同一细胞系世代项目。
结果
鉴定到差异表达 (DE) 的 miRNA，这些 miRNA 预测靶向与蛋白质折叠相关的几个蛋白质。发现 miR-200a 具有多个与未折叠蛋白反应 (UPR) 相关的预测靶标，这些靶标在高 Qp CDCL 中表达降低，并且在 mRNA 水平上没有检测到变化。在 CHO CDCL 中转染 miR-200a 可使重组蛋白滴度提高 1.2 倍，Qp 提高 1.8 倍。
结论
这些结果可能表明 miR-200a 在 UPR 的转录后调节中起作用，表明 miR-200a 是工程化具有工业吸引力的 CHO 细胞表型的潜在靶标。
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