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Endogenous p21-Dependent Transgene Control for CHO Cell Engineering
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dc.contributor.authorLee, Youngsik-
dc.contributor.authorKwak, Jin Myeong-
dc.contributor.authorLee, Jae Seong-
dc.date.issued2020-07-17-
dc.identifier.issn2161-5063-
dc.identifier.urihttps://aurora.ajou.ac.kr/handle/2018.oak/31398-
dc.identifier.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85087418073&origin=inward-
dc.description.abstractNumerous engineering efforts have been made in Chinese hamster ovary (CHO) cells for high level production of therapeutic proteins. However, the dynamic regulation of transgene expression is limited in current systems. Here, we investigated the effective regulation of transgene expression in CHO cells via targeted integration-based endogenous gene tagging with engineering target genes. Targeted integration of EGFP-human Bcl-2 into the p21 locus effectively reduced the apoptosis, compared with random populations in which Bcl-2 expression was driven by cytomegalovirus (CMV) promoter. Endogenous p21 and EGFP-human Bcl-2 displayed similar expression dynamics in batch cultures, and the antiapoptotic effect altered the expression pattern of endogenous p21 showing the mutual influences between expression of p21 and Bcl-2. We further demonstrated the inducible transgene expression by adding low concentrations of hydroxyurea. The present engineering strategy will provide a valuable CHO cell engineering tool that can be used to control dynamic transgene expression in accordance with cellular states.-
dc.description.sponsorshipThis work was supported by the NRF funded by the Korean government (2018R1C1B6001423 and 2019R1A6A1A11051471).-
dc.language.isoeng-
dc.publisherAmerican Chemical Society-
dc.subject.meshAnimals-
dc.subject.meshCell Engineering-
dc.subject.meshCHO Cells-
dc.subject.meshCricetinae-
dc.subject.meshCricetulus-
dc.subject.meshCyclin-Dependent Kinase Inhibitor p21-
dc.subject.meshGene Expression-
dc.subject.meshGreen Fluorescent Proteins-
dc.subject.meshHydroxyurea-
dc.subject.meshPromoter Regions, Genetic-
dc.subject.meshProto-Oncogene Proteins c-bcl-2-
dc.subject.meshTransgenes-
dc.titleEndogenous p21-Dependent Transgene Control for CHO Cell Engineering-
dc.typeArticle-
dc.citation.endPage1580-
dc.citation.number7-
dc.citation.startPage1572-
dc.citation.titleACS Synthetic Biology-
dc.citation.volume9-
dc.identifier.bibliographicCitationACS Synthetic Biology, Vol.9 No.7, pp.1572-1580-
dc.identifier.doi10.1021/acssynbio.9b00526-
dc.identifier.pmid32539343-
dc.identifier.scopusid2-s2.0-85087418073-
dc.identifier.urlhttp://pubs.acs.org/journal/asbcd6-
dc.subject.keywordapoptosis-
dc.subject.keywordChinese hamster ovary cells-
dc.subject.keywordendogenous gene tagging-
dc.subject.keywordp21-
dc.subject.keywordtargeted integration-
dc.subject.keywordtransgene expression regulation-
dc.type.otherArticle-
dc.description.isoafalse-
dc.subject.subareaBiomedical Engineering-
dc.subject.subareaBiochemistry, Genetics and Molecular Biology (miscellaneous)-
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College of Bio-convergence Engineering
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