Abstract:
BACKGROUND:Methylated CpG dinucleotides in promoters are associated with the loss of gene expression in recombinant Chinese hamster ovary (CHO) cells during large-scale commercial manufacturing. We evaluated a promoter devoid of CpG dinucleotides, CpGfree, in parallel with a similar CpG containing promoter, CpGrich, for their ability to maintain the expression of recombinant enhanced green fluorescent protein (EGFP) after 8 weeks of culturing. RESULTS:While the promoters gave similar transient expression levels, CpGfree clones had significantly higher average stable expression possibly due to increased resistance to early silencing during integration into the chromosome. A greater proportion of cells in clones generated using the CpGfree promoter were still expressing detectable levels of EGFP after 8 weeks but the relative expression levels measured at week 8 to those measured at week 0 did not improve compared to clones generated using the CpGrich promoter. Chromatin immunoprecipitation assays indicated that the repression of the CpGfree promoter was likely linked to histone deacetylation and methylation. Use of histone deacetylase inhibitors also managed to recover some of the lost expression. CONCLUSION:Using a promoter without CpG dinucleotides could mitigate the early gene silencing but did not improve longer-term expression stability as silencing due to histone modifications could still take place. The results presented here would aid in promoter selection and design for improved protein production in CHO and other mammalian cells.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Ho SC,Koh EY,Soo BP,Mariati,Chao SH,Yang Ydoi
10.1186/s12896-016-0300-ysubject
Has Abstractpub_date
2016-10-18 00:00:00pages
71issue
1issn
1472-6750pii
10.1186/s12896-016-0300-yjournal_volume
16pub_type
杂志文章abstract:BACKGROUND:Xenoantigens are a major source of concern with regard to the success of interspecific xenografts. GGTA1 encodes α1,3-galactosyltransferase, which is essential for the biosynthesis of galactosyl-alpha 1,3-galactose, the major xenoantigen causing hyperacute rejection. GGTA1-modified pigs, therefore, are promi...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-020-00638-7
更新日期:2020-08-18 00:00:00
abstract:BACKGROUND:Protein glycosylation is of fundamental importance in many biological systems. The discovery of N-glycosylation in bacteria and the functional expression of the N-oligosaccharyltransferase PglB of Campylobacter jejuni in Escherichia coli enabled the production of engineered glycoproteins and the study of the...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-67
更新日期:2012-09-24 00:00:00
abstract:BACKGROUND:Quantum dots (QDs) have been considered as a new and efficient probe for labeling cells non-invasively in vitro and in vivo, but fairly little is known about how QDs are eliminated from cells after labeling. The purpose of this study is to investigate the metabolism of QDs in different type of cells. RESULT...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-36
更新日期:2010-05-06 00:00:00
abstract:BACKGROUND:In a previous study, anthocyanin levels in potato plants were increased by manipulating genes connected with the flavonoid biosynthesis pathway. However, starch content and tuber yield were dramatically reduced in the transgenic plants, which over-expressed dihydroflavonol reductase (DFR). RESULTS:Transgeni...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-8-25
更新日期:2008-03-05 00:00:00
abstract:BACKGROUND:Gene therapy continues to hold great potential for treating many different types of disease and dysfunction. Safe and efficient techniques for gene transfer and expression in vivo are needed to enable gene therapeutic strategies to be effective in patients. Currently, the most commonly used methods employ re...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-62
更新日期:2011-06-02 00:00:00
abstract:BACKGROUND:Cost-effective production of industrially important enzymes is a key for their successful exploitation on industrial scale. Keeping in view the extensive industrial applications of lignin peroxidase (LiP), this study was performed to purify and characterize the LiP from an indigenous strain of Trametes versi...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-46
更新日期:2012-08-03 00:00:00
abstract:BACKGROUND:There is an increasing need to understand cell-cell interactions for cell and tissue engineering purposes, such as optimizing cell sheet constructs, as well as for examining adhesion defect diseases. For cell-sheet engineering, one major obstacle to sheet function is that cell sheets in suspension are fragil...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-17
更新日期:2013-02-26 00:00:00
abstract:BACKGROUND:Equine chorionic gonadotropin (eCG) induces super-ovulation in laboratory animals. Notwithstanding its extensive usage, limited information is available regarding the differences between the in vivo effects of natural eCG (N-eCG) and recombinant eCG (R-eCG). This study aimed to investigate the gene expressio...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-020-00653-8
更新日期:2020-11-11 00:00:00
abstract:BACKGROUND:The CRISPR/Cas9 system is increasingly used for gene inactivation in mouse zygotes, but homology-directed mutagenesis and use of inbred embryos are less established. In particular, Rosa26 knock-in alleles for the insertion of transgenes in a genomic 'safe harbor' site, have not been produced. Here we applied...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0234-4
更新日期:2016-01-16 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0259-8
更新日期:2016-03-11 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0153-9
更新日期:2015-05-28 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0342-9
更新日期:2017-02-27 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-019-0551-5
更新日期:2019-08-02 00:00:00
abstract:BACKGROUND:Domesticated einkorn (Triticum monococcum L.) is one of the oldest cultivated cereal crops in the world. Its small genome size (~ 5.7 GB), low ploidy (2n = 2x = 14, AmAm) and high genetic polymorphism make this species very attractive for use as a diploid model for understanding the genomics and proteomics o...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0477-3
更新日期:2018-10-23 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-018-0421-6
更新日期:2018-03-16 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-14
更新日期:2011-02-07 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-6-14
更新日期:2006-03-06 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-13-47
更新日期:2013-06-01 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-017-0353-6
更新日期:2017-03-24 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-11-6
更新日期:2011-01-14 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-14
更新日期:2012-04-26 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-52
更新日期:2007-08-27 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-015-0145-9
更新日期:2015-06-02 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-14-81
更新日期:2014-09-03 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-66
更新日期:2010-09-09 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-12-55
更新日期:2012-08-23 00:00:00
abstract:BACKGROUND:The detection of unknown mutations is important in research and medicine. For this purpose, a mismatch-specific endonuclease CEL I from celery has been established as a useful tool in high throughput projects. Previously, CEL I-like activities were described only in a variety of plants and could not be expre...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-7-29
更新日期:2007-06-01 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-10-13
更新日期:2010-02-17 00:00:00
abstract:BACKGROUND:Polymerase chain reaction (PCR) is one of the most important developments in modern biotechnology. However, PCR is known to introduce biases, especially during multiplex reactions. Recent studies have implicated the DNA polymerase as the primary source of bias, particularly initiation of polymerization on th...
journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/1472-6750-14-10
更新日期:2014-01-30 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
doi:10.1186/s12896-016-0235-3
更新日期:2016-01-22 00:00:00