Awakening dormant glycosyltransferases in CHO cells with CRISPRa

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...

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Bibliographic Details
Published in:Biotechnology and bioengineering 2020-02, Vol.117 (2), p.593-598
Main Authors: 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.
Format: Article
Language:English
Subjects:
Animals
Biopharmaceuticals
CHO
CHO Cells
Cricetinae
Cricetulus
CRISPR
CRISPR-Cas Systems - genetics
CRISPRa
Gene Editing - methods
Glycan
Glycosylation
Glycosyltransferases - genetics
Mgat3
Phenotype
Phenotypes
Polysaccharides - analysis
Polysaccharides - chemistry
St6gal1
Toxicity
Transcription activation
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Summary: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. Chinese hamster ovary (CHO) cells are the preferred workhorse for the biopharmaceutical industry. Glycosylation is a critical quality attribute of biopharmaceuticals, but CHO glycans differ from their human counterparts. Here we demonstrate the use of CRISPR activation (CRISPRa) to increase human‐like glycosylation in CHO cells.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.27199