Journal article
Minimizing Clonal Variation during Mammalian Cell Line Engineering for Improved Systems Biology Data Generation
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
CHO Cell Line Engineering and Design, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
Quantitative Modeling of Cell Metabolism, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
University of California at San Diego5
Department of Biotechnology and Biomedicine, Technical University of Denmark6
Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark7
Network Engineering of Eukaryotic Cell factories, Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark8
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.
| Language: | English |
|---|---|
| Publisher: | American Chemical Society |
| Year: | 2018 |
| Pages: | 2148-2159 |
| ISSN: | 21615063 |
| Types: | Journal article |
| DOI: | 10.1021/acssynbio.8b00140 |
| ORCIDs: | Grav, Lise Marie , Sergeeva, Daria , Lee, Jae Seong , Marín de Mas, Igor , Andersen, Mikael Rørdam , Nielsen, Lars Keld and Kildegaard, Helene Faustrup |
CRISPR/Cas9 Clonal variation Mammalian cells Recombinase-mediated cassette exchange Targeted integration Transcriptome
Animals CHO Cells CRISPR-Cas Systems Cell Engineering Cricetinae Cricetulus Erythropoietin Plasmids Recombinant Proteins Systems Biology Transcription, Genetic clonal variation mammalian cells recombinase-mediated cassette exchange targeted integration targetedintegration transcriptome
