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

doi:10.1002/bit.27199

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

Novo Nordisk Foundation Center for Biosustainability

Research output: Contribution to journal › Journal article › Research › peer-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 language	English
Journal	Biotechnology and Bioengineering
Volume	117
Issue number	2
Pages (from-to)	593-598
ISSN	0006-3592
DOIs	https://doi.org/10.1002/bit.27199
Publication status	Published - 2020

Keywords

CHO
CRISPRa
Glycosylation
Mgat3
St6gal11

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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.",

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AU - Julie la Cour Karottki, Karen

AU - Hefzi, Hooman

AU - Xiong, Kai

AU - Shamie, Isaac

AU - Hansen, Anders Holmgaard

AU - Li, Songyuan

AU - Pedersen, Lasse Ebdrup

AU - Li, Shangzhong

AU - Lee, Jae Seong

AU - Min Lee, Gyun

AU - Kildegaard, Helene Faustrup

AU - Lewis, Nathan E.

PY - 2020

Y1 - 2020

N2 - 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.

AB - 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.

KW - CHO

KW - CRISPRa

KW - Glycosylation

KW - Mgat3

KW - St6gal11

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DO - 10.1002/bit.27199

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JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

IS - 2

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Awakening dormant glycosyltransferases in CHO cells with CRISPRa

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