Identification of consensus sequence from matrix attachment regions and functional analysis of its activity in stably transfected Chinese hamster ovary cells

Matrix attachment regions (MARs) can enhance transgene expression levels and maintain stability. However, the consensus sequence from MARs and its functional analysis remains to be examined. Here, we assessed a possible consensus sequence from MARs and assessed its activity in stably transfected Chi...

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Bibliographic Details
Published in:Journal of cellular biochemistry 2019-08, Vol.120 (8), p.13985-13993
Main Authors: Gao, Jian‐Hui, Wang, Tian‐Yun, Zhang, Mao‐Ying, Shi, Fang, Gu, Shan‐Zhi
Format: Article
Language:English
Subjects:
Bioinformatics
Chinese hamster ovary cell
consensus sequence
Conserved sequence
Downstream
Flow cytometry
Flow stability
Fluorescence
Functional analysis
Green fluorescent protein
matrix attachment region
Matrix attachment regions
Nucleotide sequence
Stability analysis
Toxicity
transgene expression
Upstream
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Summary:Matrix attachment regions (MARs) can enhance transgene expression levels and maintain stability. However, the consensus sequence from MARs and its functional analysis remains to be examined. Here, we assessed a possible consensus sequence from MARs and assessed its activity in stably transfected Chinese hamster ovary (CHO) cells. First, we analyzed the effects of 10 MARs on transfected CHO cells and then analyzed the consensus motifs from these MARs using a bioinformatics method. The consensus sequence was synthesized and cloned upstream or downstream of the eukaryotic vector. The constructs were transfected into CHO cells and the expression levels and stability of enhanced green fluorescent protein were detected by flow cytometry. The results indicated that eight of the ten MARs increased transgene expression in transfected CHO cells. Three consensus motifs were found after bioinformatics analyses. The consensus sequence tandemly enhanced transgene expression when it was inserted into the eukaryotic expression vector; the effect of the addition upstream was stronger than that downstream. Thus, we found a MAR consensus sequence that may regulate the MAR‐mediated increase in transgene expression. Assessed the possible consensus sequence from matrix attachment regions (MARs) and assessed its activity in stably transfected Chinese hamster ovary cells. First found the MAR consensus sequence that may mediate the MAR activity of increasing transgene expression. The levels of transgene expression were not only copy‐number‐dependent, other mechanism may be involved.
ISSN:0730-2312
1097-4644
DOI:10.1002/jcb.28673