Reduced apoptosis in Chinese hamster ovary cells via optimized CRISPR interference

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Chinese hamster ovary (CHO) cells are widely used for biopharmaceutical protein production. One challenge limiting CHO cell productivity is apoptosis stemming from cellular stress during protein production. Here we applied CRISPR interference (CRISPRi) to downregulate the endogenous expression of apoptotic genes Bak, Bax, and Casp3 in CHO cells. In addition to reduced apoptosis, mitochondrial membrane integrity was improved and the caspase activity was reduced. Moreover, we optimized the CRISPRi system to enhance the gene repression efficiency in CHO cells by testing different repressor fusion types. An improved Cas9 repressor has been identified by applying C-terminal fusion of a bipartite repressor domain, KRAB-MeCP2, to nuclease-deficient Cas9. These results collectively demonstrate that CHO cells can be rescued from cell apoptosis by targeted gene repression using the CRISPRi system.
Publisher
WILEY
Issue Date
2019-06
Language
English
Article Type
Article
Citation

BIOTECHNOLOGY AND BIOENGINEERING, v.116, no.7, pp.1813 - 1819

ISSN
0006-3592
DOI
10.1002/bit.26969
URI
http://hdl.handle.net/10203/262788
Appears in Collection
BS-Journal Papers(저널논문)
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