title: Ribosome profiling-guided depletion of an mRNA increases cell growth rate and protein secretion.
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.
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors.forename: Thomas Beuchert
authors.initials: TB
authors.lastname: Kallehauge
authors.affiliation: Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, United States.|Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA 92093, United States.
authors.forename: Shangzhong
authors.initials: S
authors.lastname: Li
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors.forename: Lasse Ebdrup
authors.initials: LE
authors.lastname: Pedersen
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors.forename: Tae Kwang
authors.initials: TK
authors.lastname: Ha
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.|Department of Systems Biology, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors.forename: Daniel
authors.initials: D
authors.lastname: Ley
authors.affiliation: Department of Systems Biology, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors.forename: Mikael Rørdam
authors.initials: MR
authors.lastname: Andersen
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors.forename: Helene Faustrup
authors.initials: HF
authors.lastname: Kildegaard
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.|Department of Biological Sciences, KAIST, Daejeon 305-701, Korea.
authors.forename: Gyun Min
authors.initials: GM
authors.lastname: Lee
authors.affiliation: Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA 92093, United States.|Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, CA 92093, United States.
authors.forename: Nathan E
authors.initials: NE
authors.lastname: Lewis
mesh_terms: D000818:Animals; D016466:CHO Cells; D002452:Cell Count; D049109:Cell Proliferation / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D055785:Gene Knockdown Techniques; D007074:Immunoglobulin G / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D007074:Immunoglobulin G; D009711:Nucleotides; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins