Datasets

Title	Ribosome profiling-guided depletion of an mRNA improves CHO cell growth and recombinant product titers
Organism
Number of Samples	12
Release Date	2016/03/23 00:00
Sequencing Types
Protocol Details

Title
Authors	Kallehauge TB,Li S,Pedersen LE,Ha TK,Ley D,Andersen MR,Kildegaard HF,Lee GM,Lewis NE
Journal	Scientific reports
Publication Date	2017 Jan 16
Abstract	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.
PMC	PMC5238448
PMID	28091612
DOI

Kallehauge et al. 2016 (PRJNA316065)