DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Che Lin | ko |
dc.contributor.author | Lee, Gyun Min | ko |
dc.date.accessioned | 2018-12-20T08:00:00Z | - |
dc.date.available | 2018-12-20T08:00:00Z | - |
dc.date.created | 2018-11-26 | - |
dc.date.created | 2018-11-26 | - |
dc.date.created | 2018-11-26 | - |
dc.date.issued | 2019-03 | - |
dc.identifier.citation | METABOLIC ENGINEERING, v.52, pp.57 - 67 | - |
dc.identifier.issn | 1096-7176 | - |
dc.identifier.uri | http://hdl.handle.net/10203/248654 | - |
dc.description.abstract | A Chinese hamster ovary (CHO) cell line producing recombinant human bone morphogenetic protein-4 (rhBMP-4) (CHO-BMP-4), which expresses essential components of BMP signal transduction, underwent autocrine BMP-4 signaling. RNA seq analysis on CHO host cells (DG44) treated with rhBMP-4 (20 µg/mL) suggested that rhBMP-4 induced signaling in CHO cells could be a critical factor in limiting rhBMP-4 production and should be removed to improve rhBMP-4 production in recombinant CHO (rCHO) cells. The inhibition of autocrine BMP signaling in CHO-BMP-4 cells by the addition of LDN-193189, a chemical inhibitor of BMP receptor type I, significantly increased the mRNA expression levels of rhBMP-4. To establish BMP signaling-free host cells, a BMP receptor, the BMPRIA or BMPRII gene in DG44 cells, was knocked out using CRISPR/Cas9 gene-editing technology. Using three different knockout (KO) host cell lines as well as a DG44 wild-type (wt) cell line, rCHO cell clones producing rhBMP-4 were generated by a stepwise selection with increasing methotrexate concentrations. KO-derived clones showed a significantly higher maximum rhBMP-4 concentration than wt-derived clones in both batch and fed-batch cultures. Unlike wt-derived clones, KO-derived cell clones were able to produce higher amounts of hBMP-4 transcripts and proteins in the stationary phase of growth and did not experience growth inhibition induced by rhBMP-4. The mean maximum rhBMP-4 concentration of KO host-derived clones was approximately 2.4-fold higher than that of wt-derived clones (P < 0.05). Taken together, the disruption of BMP signaling in CHO cells by knocking out the BMP receptor significantly improved rhBMP-4 production. | - |
dc.language | English | - |
dc.publisher | ACADEMIC PRESS INC ELSEVIER SCIENCE | - |
dc.title | Improving recombinant bone morphogenetic protein-4 (BMP-4) production by autoregulatory feedback loop removal using BMP receptor-knockout CHO cell lines | - |
dc.type | Article | - |
dc.identifier.wosid | 000457633200006 | - |
dc.identifier.scopusid | 2-s2.0-85056823326 | - |
dc.type.rims | ART | - |
dc.citation.volume | 52 | - |
dc.citation.beginningpage | 57 | - |
dc.citation.endingpage | 67 | - |
dc.citation.publicationname | METABOLIC ENGINEERING | - |
dc.identifier.doi | 10.1016/j.ymben.2018.11.003 | - |
dc.contributor.localauthor | Lee, Gyun Min | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Bone morphogenetic protein-4 (BMP-4) | - |
dc.subject.keywordAuthor | BMP signaling | - |
dc.subject.keywordAuthor | CHO cells | - |
dc.subject.keywordAuthor | BMP receptor knockout | - |
dc.subject.keywordAuthor | CRISPR/Cas9 | - |
dc.subject.keywordPlus | HAMSTER OVARY CELLS | - |
dc.subject.keywordPlus | GENE-EXPRESSION | - |
dc.subject.keywordPlus | APOPTOSIS | - |
dc.subject.keywordPlus | DIFFERENTIATION | - |
dc.subject.keywordPlus | CARCINOMA | - |
dc.subject.keywordPlus | DELETION | - |
dc.subject.keywordPlus | ADAM8 | - |
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