Native Transcriptome Profiling of Virus-Infected Chinese Hamster Ovary Cells
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Issue Date
2021-12
Authors
Sher, Melissa '21
Degree
MS in Molecular Biosciences
Advisor
Yager, Eric
Committee Members
Singh, Vir
VanHoute, David
Kelliher, Jennifer
VanHoute, David
Kelliher, Jennifer
Journal Title
Journal ISSN
Volume Title
Abstract
Chinese Hamster Ovary (CHO) cells, used extensively to produce therapeutic recombinant proteins, may become infected with viruses without apparent impact to cell metabolism or morphology. As contamination is currently detected using lengthy culture-based methods, (Pharmacopeia U. S., USP Viral Safety of Biotechnology Products Derived from Cell Lines of Human or Animal Origin ) (Pharmacopeia U. S., Chapter Virology Test Methods, Current Version) contamination may spread before an infection is discovered, (RL, 1998) thereby squandering resources and impacting the amount of therapeutics available to patients. A molecular diagnostic method quantifying genes known to be differentially expressed (DEGs) during viral infection has the potential to facilitate early identification of viral contaminants. Unfortunately, the transcriptional landscape of CHO cells during infection is not well documented, and the proposed diagnostic method cannot be designed without thorough characterization of healthy and infected CHO transcriptomes. The present study characterized the transcriptome of CHO cells (healthy and infected with a common viral contaminant, Minute Virus of Mice, (MVM) (RL, 1998) (Jacoby RO, 2003)) whose mechanism of infection is known to impact gene expression (Adeyemi RO P. D., 2014) (Majumder K, 2017) using native RNA nanopore sequencing (Oxford Nanopore Technologies). Native transcriptome analysis of virus-infected CHO cells elucidated a panel of detectable genes which were differentially expressed during viral infection. Differential expression of these biomarkers could not be confirmed by an orthogonal RT-PCR. However, alternative uses of native transcriptome sequencing are promising for development of a rapid, non-specific molecular diagnostic method to monitor the health of CHO cell cultures.
Citation
Sher M. Native transcriptome profiling of virus-infected Chinese hamster ovary cells [thesis]. Ann Arbor (MI): Proquest LLC; 2021. 76 p.
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