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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.
Afiliación
  • Grav LM; The Novo Nordisk Foundation Center for Biosustainability , Technical University of Denmark , 2800 Kgs. Lyngby , Denmark.
  • Sergeeva D; The Novo Nordisk Foundation Center for Biosustainability , Technical University of Denmark , 2800 Kgs. Lyngby , Denmark.
  • Lee JS; The Novo Nordisk Foundation Center for Biosustainability , Technical University of Denmark , 2800 Kgs. Lyngby , Denmark.
  • Marin de Mas I; Department of Molecular Science and Technology , Ajou University , Suwon 16499 , Republic of Korea.
  • Lewis NE; The Novo Nordisk Foundation Center for Biosustainability , Technical University of Denmark , 2800 Kgs. Lyngby , Denmark.
  • Andersen MR; Department of Pediatrics , University of California, San Diego , La Jolla , California 92093 , United States.
  • Nielsen LK; The Novo Nordisk Foundation Center for Biosustatainability , University of California, San Diego , La Jolla , California 92093 , United States.
  • Lee GM; Department of Biotechnology and Biomedicine , Technical University of Denmark , 2800 Kgs. Lyngby , Denmark.
  • Kildegaard HF; The Novo Nordisk Foundation Center for Biosustainability , Technical University of Denmark , 2800 Kgs. Lyngby , Denmark.
ACS Synth Biol ; 7(9): 2148-2159, 2018 09 21.
Article en En | MEDLINE | ID: mdl-30060646
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.
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Texto completo: 1 Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Biología de Sistemas / Ingeniería Celular Límite: Animals Idioma: En Revista: ACS Synth Biol Año: 2018 Tipo del documento: Article País de afiliación: Dinamarca

Texto completo: 1 Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Biología de Sistemas / Ingeniería Celular Límite: Animals Idioma: En Revista: ACS Synth Biol Año: 2018 Tipo del documento: Article País de afiliación: Dinamarca