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Minimizing Clonal Variation during Mammalian Cell Line Engineering for Improved Systems Biology Data Generation
  • Grav, Lise Marie ;
  • Sergeeva, Daria ;
  • Lee, Jae Seong ;
  • Marin De Mas, Igor ;
  • Lewis, Nathan E. ;
  • Andersen, Mikael Rørdam ;
  • Nielsen, Lars Keld ;
  • Lee, Gyun Min ;
  • Kildegaard, Helene Faustrup
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Publication Year
2018-09-21
Journal
ACS Synthetic Biology
Publisher
American Chemical Society
Citation
ACS Synthetic Biology, Vol.7 No.9, pp.2148-2159
Keyword
clonal variationCRISPR/Cas9mammalian cellsrecombinase-mediated cassette exchangetargeted integrationtranscriptome
Mesh Keyword
AnimalsCell EngineeringCHO CellsCricetinaeCricetulusCRISPR-Cas SystemsErythropoietinPlasmidsRecombinant ProteinsSystems BiologyTranscription, GeneticTranscriptome
All Science Classification Codes (ASJC)
Biomedical EngineeringBiochemistry, Genetics and Molecular Biology (miscellaneous)
Abstract
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.
ISSN
2161-5063
Language
eng
URI
https://aurora.ajou.ac.kr/handle/2018.oak/30337
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85052316550&origin=inward
DOI
https://doi.org/2-s2.0-85052316550
Journal URL
http://pubs.acs.org/journal/asbcd6
Type
Article
Funding
The authors thank Nachon Charanyanonda Petersen and Karen Kathrine Br\u00f8ndum for assistance with the FACS. We greatly acknowledge support from the Novo Nordisk Foundation (NNF10CC1016517 and NNF16CC0020908).
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College of Bio-convergence Engineering
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