Minimizing Clonal Variation during Mammalian Cell Line Engineering for Improved Systems Biology Data Generation

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Mammalian cells are widely used to express genes for basic biology studies and biopharmaceuticals. Current methods for generation of engineered cell lines introduce high genomic and phenotypic diversity, which hamper studies of gene functions and discovery of novel cellular mechanisms. Here, we minimized clonal variation by integrating a landing pad for recombinase-mediated cassette exchange site-specifically into the genome of CHO cells using CRISPR and generated subclones expressing four different recombinant proteins. The subclones showed low clonal variation with high consistency in growth, transgene transcript levels and global transcriptional response to recombinant protein expression, enabling improved studies of the impact of transgenes on the host transcriptome. Little variation over time in subclone phenotypes and transcriptomes was observed when controlling environmental culture conditions. The platform enables robust comparative studies of genome engineered CHO cell lines and can be applied to other mammalian cells for diverse biological, biomedical and biotechnological applications.
Publisher
AMER CHEMICAL SOC
Issue Date
2018-09
Language
English
Article Type
Article
Keywords

MEDIATED CASSETTE EXCHANGE; SITE-SPECIFIC INTEGRATION; HAMSTER OVARY CELLS; CHO-CELLS; ANTIBODY EXPRESSION; QUANTIFICATION; ERYTHROPOIETIN; CRISPR/CAS9; PLATFORM; PATHWAY

Citation

ACS SYNTHETIC BIOLOGY, v.7, no.9, pp.2148 - 2159

ISSN
2161-5063
DOI
10.1021/acssynbio.8b00140
URI
http://hdl.handle.net/10203/246218
Appears in Collection
BS-Journal Papers(저널논문)
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