Translatome analysis of CHO cells to identify key growth genes

Autor: Sze Wai Ng, Hsueh Lee Lim, Bernard Loo, Franck C. Courtes, Miranda G.S. Yap, Niki S.C. Wong, Leah A. Vardy, Geoffrey Koh, Joyce Lin, Dong-Yup Lee
Rok vydání: 2013
Předmět:
Zdroj: Journal of Biotechnology. 167:215-224
ISSN: 0168-1656
DOI: 10.1016/j.jbiotec.2013.07.010
Popis: We report the first investigation of translational efficiency on a global scale, also known as translatome, of a Chinese hamster ovary (CHO) DG44 cell line producing monoclonal antibodies (mAb). The translatome data was generated via combined use of high resolution and streamlined polysome profiling technology and proprietary Nimblegen microarrays probing for more than 13K annotated CHO-specific genes. The distribution of ribosome loading during the exponential growth phase revealed the translational activity corresponding to the maximal growth rate, thus allowing us to identify stably and highly translated genes encoding heterogeneous nuclear ribonucleoproteins (Hnrnpc and Hnrnpa2b1), protein regulator of cytokinesis 1 (Prc1), glucose-6-phosphate dehydrogenase (G6pdh), UTP6 small subunit processome (Utp6) and RuvB-like protein 1 (Ruvbl1) as potential key players for cellular growth. Moreover, correlation analysis between transcriptome and translatome data sets showed that transcript level and translation efficiency were uncoupled for 95% of investigated genes, suggesting the implication of translational control mechanisms such as the mTOR pathway. Thus, the current translatome analysis platform offers new insights into gene expression in CHO cell cultures by bridging the gap between transcriptome and proteome data, which will enable researchers of the bioprocessing field to prioritize in high-potential candidate genes and to devise optimal strategies for cell engineering toward improving culture performance.
Databáze: OpenAIRE
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