Ribosome profiling-guided depletion of an mRNA increases cell growth rate and protein secretion

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Recombinant protein production coopts the host cell machinery to provide high protein yields of industrial enzymes or biotherapeutics. However, since protein translation is energetically expensive and tightly controlled, it is unclear if highly expressed recombinant genes are translated as efficiently as host genes. Furthermore, it is unclear how the high expression impacts global translation. Here, we present the first genome-wide view of protein translation in an IgG-producing CHO cell line, measured with ribosome profiling. Through this we found that our recombinant mRNAs were translated as efficiently as the host cell transcriptome, and sequestered up to 15% of the total ribosome occupancy. During cell culture, changes in recombinant mRNA translation were consistent with changes in transcription, demonstrating that transcript levels influence specific productivity. Using this information, we identified the unnecessary resistance marker NeoR to be a highly transcribed and translated gene. Through siRNA knock-down of NeoR, we improved the production-and growth capacity of the host cell. Thus, ribosomal profiling provides valuable insights into translation in CHO cells and can guide efforts to enhance protein production.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2017-01
Language
English
Article Type
Article
Keywords

HAMSTER OVARY CELLS; CHINESE-HAMSTER; CHO-CELLS; TRANSLATIONAL CONTROL; MTOR PATHWAY; IN-VIVO; EXPRESSION; GENOME; GENES; RESOLUTION

Citation

SCIENTIFIC REPORTS, v.7, pp.1 - 10

ISSN
2045-2322
DOI
10.1038/srep40388
URI
http://hdl.handle.net/10203/220726
Appears in Collection
BS-Journal Papers(저널논문)
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