Awakening dormant glycosyltransferases in CHO cells with CRISPRa

Karen Julie la Cour Karottki, Hooman Hefzi, Kai Xiong, Isaac Shamie, Anders Holmgaard Hansen, Songyuan Li, Lasse Ebdrup Pedersen, Shangzhong Li, Jae Seong Lee, Gyun Min Lee, Helene Faustrup Kildegaard*, Nathan E. Lewis

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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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.
Original languageEnglish
JournalBiotechnology and Bioengineering
Volume117
Issue number2
Pages (from-to)593-598
ISSN0006-3592
DOIs
Publication statusPublished - 2020

Keywords

  • CHO
  • CRISPRa
  • Glycosylation
  • Mgat3
  • St6gal11

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