Awakening dormant glycosyltransferases in CHO cells with CRISPRa
Awakening dormant glycosyltransferases in CHO cells with CRISPRa
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DOI：
10.1002/bit.27199
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发表时间：
2019-11-12
影响因子：
3.8
通讯作者：
Lewis, Nathan E.
中科院分区：
文献类型：
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作者：
Karottki, Karen Julie la Cour;Hefzi, Hooman;Lewis, Nathan E.
中国仓鼠卵巢（CHO）细胞是生物制药行业的首选主力，CRISPR/Cas9已被证明在CHO细胞中产生靶向基因扰动方面具有强大的作用。在这里，我们用CRISPR激活（CRISPRa）扩展了CRISPR工程工具箱，以增加内源基因的转录。我们成功地增加了Mgat 3和St 6 gal 1的转录，并通过随后检测产生的适当聚糖结构在功能水平上验证了它们的活性。这项研究表明，CRISPRa可以针对所需表型对CHO细胞进行靶向改变。
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.