DC Field | Value | Language |
---|---|---|
dc.contributor.author | Lee, So-Jeong | ko |
dc.contributor.author | Lee, Gyun-Min | ko |
dc.date.accessioned | 2015-01-27T05:09:47Z | - |
dc.date.available | 2015-01-27T05:09:47Z | - |
dc.date.created | 2013-07-18 | - |
dc.date.created | 2013-07-18 | - |
dc.date.issued | 2013-09 | - |
dc.identifier.citation | PROCESS BIOCHEMISTRY, v.48, no.9, pp.1436 - 1440 | - |
dc.identifier.issn | 1359-5113 | - |
dc.identifier.uri | http://hdl.handle.net/10203/193182 | - |
dc.description.abstract | Transient gene expression (TGE) in Chinese hamster ovary (CHO) cells with polyethylenimine (PEI) as a transfection reagent has been considered as an attractive method to produce recombinant proteins rapidly for pre-clinical studies. A high level of transfection efficiency, which is required for high-level TGE in CHO cells, can be achieved by increasing the PEI concentration. However, PEI induces cytotoxicity in a dose-dependent manner. To overcome this problem, Bcl-2 protein, an anti-apoptotic protein, was overexpressed in CHO cells (DG44). At a ratio of PEI to DNA (an N/P ratio) of 10, there were no significant differences in transfection efficiency and cell viability between Bcl-2 overexpressing and non-overexpressing cells. The transfection efficiency and cell viability were 2-11% and 83-92%, respectively. However, there were significant differences (P<0.05) in the transfection efficiency and cell viability between them at a higher N/P ratio. At an NIP ratio of 40, the transfection efficiency and cell viability of Bcl-2 non-overexpressing cells were 24-38% and 35-40%, respectively, while those of Bcl-2 overexpressing cells were 48-53% and 43-56%, respectively. Furthermore, compared with Bcl-2 non-overexpressing cells, more DNAs entered the Bcl-2 overexpressing cells, resulting in a higher rate of TGE per cell. PE-Annexin V apoptosis revealed that Bcl-2 overexpression suppressed PEI-induced apoptotic cell death at high N/P ratios. Taken together, Bcl-2 overexpression in CHO cells suppresses apoptotic cell death during PEI-mediated transient transfection, resulting in enhanced transfection efficiency and TGE. (C) 2013 Elsevier Ltd. All rights reserved. | - |
dc.language | English | - |
dc.publisher | ELSEVIER SCI LTD | - |
dc.subject | HAMSTER OVARY CELLS | - |
dc.subject | PROTEIN EXPRESSION | - |
dc.subject | TRANSIENT TRANSFECTION | - |
dc.subject | EBNA1 CELLS | - |
dc.subject | CULTURE | - |
dc.subject | ENHANCEMENT | - |
dc.subject | SUSPENSION | - |
dc.subject | DEATH | - |
dc.subject | CYTOTOXICITY | - |
dc.subject | ANTIBODIES | - |
dc.title | Bcl-2 overexpression in CHO cells improves polyethylenimine-mediated gene transfection | - |
dc.type | Article | - |
dc.identifier.wosid | 000324349500024 | - |
dc.identifier.scopusid | 2-s2.0-84882916291 | - |
dc.type.rims | ART | - |
dc.citation.volume | 48 | - |
dc.citation.issue | 9 | - |
dc.citation.beginningpage | 1436 | - |
dc.citation.endingpage | 1440 | - |
dc.citation.publicationname | PROCESS BIOCHEMISTRY | - |
dc.identifier.doi | 10.1016/j.procbio.2013.03.021 | - |
dc.contributor.localauthor | Lee, Gyun-Min | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | CHO | - |
dc.subject.keywordAuthor | Bcl-2 | - |
dc.subject.keywordAuthor | PEI | - |
dc.subject.keywordAuthor | Transient gene expression | - |
dc.subject.keywordAuthor | Apoptosis | - |
dc.subject.keywordPlus | HAMSTER OVARY CELLS | - |
dc.subject.keywordPlus | PROTEIN EXPRESSION | - |
dc.subject.keywordPlus | TRANSIENT TRANSFECTION | - |
dc.subject.keywordPlus | EBNA1 CELLS | - |
dc.subject.keywordPlus | CULTURE | - |
dc.subject.keywordPlus | ENHANCEMENT | - |
dc.subject.keywordPlus | SUSPENSION | - |
dc.subject.keywordPlus | DEATH | - |
dc.subject.keywordPlus | CYTOTOXICITY | - |
dc.subject.keywordPlus | ANTIBODIES | - |
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