Ribosome profiling-guided depletion of an mRNA increases cell growth rate and protein secretion
| Autor: | Thomas Beuchert Kallehauge, Tae Kwang Ha, Shangzhong Li, Mikael Rørdam Andersen, Lasse Ebdrup Pedersen, Nathan E. Lewis, Gyun Min Lee, Helene Faustrup Kildegaard, Daniel Ley |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Transcription Genetic Cell Survival 1.1 Normal biological development and functioning Messenger Cell Count CHO Cells Biology Ribosome Article Transcriptome 03 medical and health sciences Cricetulus Genetic Underpinning research Cricetinae Protein biosynthesis Genetics Animals Ribosome profiling RNA Messenger Cell Proliferation Messenger RNA Multidisciplinary Nucleotides Chinese hamster ovary cell Human Genome Proteins Translation (biology) Molecular biology Recombinant Proteins Cell biology 030104 developmental biology Cell culture Immunoglobulin G Protein Biosynthesis Gene Knockdown Techniques RNA Generic health relevance Infection Ribosomes Transcription Biotechnology |
| Zdroj: | Beuchert Kallehauge, T, Li, S, Pedersen, L E, Kwang Ha, T, Ley, D, Andersen, M R, Kildegaard, H F, Min Lee, G & Lewis, N E 2017, ' Ribosome profiling-guided depletion of an mRNA increases cell growth rate and protein secretion ', Scientific Reports, vol. 7, 40388 . https://doi.org/10.1038/srep40388 Scientific reports, vol 7, iss 1 Scientific Reports Beuchert Kallehauge, T, Li, S, Pedersen, L E, Kwang Ha, T, Ley, D, Andersen, M R, Kildegaard, H F, Min Lee, G & Lewis, N E 2017, ' Ribosome profiling-guided depletion of an mRNA increases cell growth rate and protein secretion ' Scientific Reports, vol 7, 40388 . DOI: 10.1038/srep40388 |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep40388 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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