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

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

Cite this

Julie la Cour Karottki, K., Hefzi, H., Xiong, K., Shamie, I., Hansen, A. H., Li, S., ... Lewis, N. E. (2020). Awakening dormant glycosyltransferases in CHO cells with CRISPRa. Biotechnology and Bioengineering, 117(2), 593-598. https://doi.org/10.1002/bit.27199
Julie la Cour Karottki, Karen ; Hefzi, Hooman ; Xiong, Kai ; Shamie, Isaac ; Hansen, Anders Holmgaard ; Li, Songyuan ; Pedersen, Lasse Ebdrup ; Li, Shangzhong ; Lee, Jae Seong ; Min Lee, Gyun ; Kildegaard, Helene Faustrup ; Lewis, Nathan E. / Awakening dormant glycosyltransferases in CHO cells with CRISPRa. In: Biotechnology and Bioengineering. 2020 ; Vol. 117, No. 2. pp. 593-598.
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keywords = "CHO, CRISPRa, Glycosylation, Mgat3, St6gal11",
author = "{Julie la Cour Karottki}, Karen and Hooman Hefzi and Kai Xiong and Isaac Shamie and Hansen, {Anders Holmgaard} and Songyuan Li and Pedersen, {Lasse Ebdrup} and Shangzhong Li and Lee, {Jae Seong} and {Min Lee}, Gyun and Kildegaard, {Helene Faustrup} and Lewis, {Nathan E.}",
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Julie la Cour Karottki, K, Hefzi, H, Xiong, K, Shamie, I, Hansen, AH, Li, S, Pedersen, LE, Li, S, Lee, JS, Min Lee, G, Kildegaard, HF & Lewis, NE 2020, 'Awakening dormant glycosyltransferases in CHO cells with CRISPRa', Biotechnology and Bioengineering, vol. 117, no. 2, pp. 593-598. https://doi.org/10.1002/bit.27199

Awakening dormant glycosyltransferases in CHO cells with CRISPRa. / Julie la Cour Karottki, Karen; Hefzi, Hooman; Xiong, Kai; Shamie, Isaac; Hansen, Anders Holmgaard; Li, Songyuan; Pedersen, Lasse Ebdrup; Li, Shangzhong; Lee, Jae Seong; Min Lee, Gyun; Kildegaard, Helene Faustrup; Lewis, Nathan E.

In: Biotechnology and Bioengineering, Vol. 117, No. 2, 2020, p. 593-598.

Research output: Contribution to journalJournal articleResearchpeer-review

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T1 - Awakening dormant glycosyltransferases in CHO cells with CRISPRa

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

U2 - 10.1002/bit.27199

DO - 10.1002/bit.27199

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EP - 598

JO - Biotechnology and Bioengineering (Print)

JF - Biotechnology and Bioengineering (Print)

SN - 0006-3592

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