Awakening dormant glycosyltransferases in CHO cells with CRISPRa.

Karen Julie la Cour KarottkiHooman HefziKai XiongIsaac ShamieAnders Holmgaard HansenSongyuan LiLasse Ebdrup Pedersen Shangzhong Li Jae Seong Lee Gyun Min Lee Helene Faustrup Kildegaard Nathan E Lewis

Published in: Biotechnology and bioengineering (2019)

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.

Keyphrases

induced apoptosis
crispr cas
cell cycle arrest
genome editing
genome wide
endoplasmic reticulum stress
oxidative stress
cell death
gene expression
pi k akt
copy number
cell proliferation