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Awakening dormant glycosyltransferases in CHO cells with CRISPRa.
Karottki, Karen Julie la Cour; Hefzi, Hooman; Xiong, Kai; Shamie, Isaac; Hansen, Anders Holmgaard; Li, Songyuan; Pedersen, Lasse Ebdrup; Li, Shangzhong; Lee, Jae Seong; Lee, Gyun Min; Kildegaard, Helene Faustrup; Lewis, Nathan E.
Afiliação
  • Karottki KJC; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
  • Hefzi H; Department of Pediatrics, University of California San Diego, San Diego, California.
  • Xiong K; The Novo Nordisk Foundation Center for Biosustainability, University of California San Diego, San Diego, California.
  • Shamie I; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
  • Hansen AH; Department of Pediatrics, University of California San Diego, San Diego, California.
  • Li S; The Novo Nordisk Foundation Center for Biosustainability, University of California San Diego, San Diego, California.
  • Pedersen LE; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
  • Li S; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
  • Lee JS; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
  • Lee GM; Department of Pediatrics, University of California San Diego, San Diego, California.
  • Kildegaard HF; The Novo Nordisk Foundation Center for Biosustainability, University of California San Diego, San Diego, California.
  • Lewis NE; Department of Bioengineering, University of California San Diego, San Diego, California.
Biotechnol Bioeng ; 117(2): 593-598, 2020 02.
Article em En | MEDLINE | ID: mdl-31631317
ABSTRACT
Chinese hamster ovary (CHO) cells are the preferred workhorse for the biopharmaceutical industry, and CRISPR/Cas9 has proven powerful for generating targeted gene perturbations in CHO cells. Here, we expand the CRISPR engineering toolbox with CRISPR activation (CRISPRa) to increase transcription of endogenous genes. We successfully increased transcription of Mgat3 and St6gal1, and verified their activity on a functional level by subsequently detecting that the appropriate glycan structures were produced. This study demonstrates that CRISPRa can make targeted alterations of CHO cells for desired phenotypes.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Glicosiltransferases / Sistemas CRISPR-Cas / Edição de Genes Idioma: En Ano de publicação: 2020 Tipo de documento: Article
Texto completo
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PubMed Links
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Glicosiltransferases / Sistemas CRISPR-Cas / Edição de Genes Idioma: En Ano de publicação: 2020 Tipo de documento: Article