University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal

Logo BOKU Resarch Portal

Selected Publication:

Maccani, A; Hackl, M; Leitner, C; Steinfellner, W; Graf, AB; Tatto, NE; Karbiener, M; Scheideler, M; Grillari, J; Mattanovich, D; Kunert, R; Borth, N; Grabherr, R; Ernst, W.
(2014): Identification of microRNAs specific for high producer CHO cell lines using steady-state cultivation
APPL MICROBIOL BIOT. 2014; 98(17): 7535-7548. FullText FullText_BOKU

Abstract:
MicroRNAs are short non-coding RNAs that play an important role in the regulation of gene expression. Hence, microRNAs are considered as potential targets for engineering of Chinese hamster ovary (CHO) cells to improve recombinant protein production. Here, we analyzed and compared the microRNA expression patterns of high, low, and non-producing recombinant CHO cell lines expressing two structurally different model proteins in order to identify microRNAs that are involved in heterologous protein synthesis and secretion and thus might be promising targets for cell engineering to increase productivity. To generate reproducible and comparable data, the cells were cultivated in a bioreactor under steady-state conditions. Global microRNA expression analysis showed that mature microRNAs were predominantly upregulated in the producing cell lines compared to the non-producer. Several microRNAs were significantly differentially expressed between high and low producers, but none of them commonly for both model proteins. The identification of target messenger RNAs (mRNAs) is essential to understand the biological function of microRNAs. Therefore, we negatively correlated microRNA and global mRNA expression data and combined them with computationally predicted and experimentally validated targets. However, statistical analysis of the identified microRNA-mRNA interactions indicated a considerable false positive rate. Our results and the comparison to published data suggest that the reaction of CHO cells to the heterologous protein expression is strongly product- and/or clone-specific. In addition, this study highlights the urgent need for reliable CHO-specific microRNA target prediction tools and experimentally validated target databases in order to facilitate functional analysis of high-throughput microRNA expression data in CHO cells.
Authors BOKU Wien:
Borth Nicole
Ernst Wolfgang
Grabherr Reingard
Grillari Johannes
Hackl Matthias
Kunert Renate
Leitner Christian
Maccani Andreas
Mattanovich Diethard
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Animals
Bioreactors
CHO Cells/physiology*
Cell Culture Techniques/methods
Cricetulus
Gene Expression Profiling
Gene Expression Regulation
MicroRNAs/metabolism*

Find related publications in this database (Keywords)
Chinese hamster ovary cells
Productivity
Chemostat
Microarray
miRNA expression profiling
miRNA target identification


Altmetric:
© BOKU Wien Imprint