Characteristic element of matrix attachment region mediates vector attachment and enhances nerve growth factor expression in Chinese hamster ovary cells

Abstract

ABSTRACT. Preliminary studies have suggested that a characteristic element of the matrix attachment region (MAR) in human interferon-β mediates the adhesion of vectors to Chinese hamster ovary (CHO) cells. In this study, we investigated if vector adhesion increased nerve growth factor (NGF) expression in CHO cells. The MAR characteristic element sequence of human interferon-β was inserted into the multiple-cloning site of the pEGFP-C1 vector. The target NGF gene was inserted upstream of the MAR characteristic element sequence to construct the MAR/ NGF expression vector. The recombinant plasmid was transfected into CHO cells and stable monoclonal cells were selected using G418. NGF mRNA and protein expression was detected by reverse transcriptasepolymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Plasmid reduction experiments were used to determine the state of transfected plasmid in mammalian cells. 9191-9199 (2015) into the vector increased NGF expression levels in CHO cells (1.93-fold) compared to the control. The recombinant plasmid expressing the MAR sequence was digested into a linear space vector. The inserted MAR and NGF sequences were consistent with those inserted into the plasmid before recombination. Therefore, we concluded that the MAR characteristic element mediates vector adhesion to CHO cells and enhances the stability and efficiency of the target gene expression.

Evaluation of the Effects of Human Beta-Interferon Scaffold Attachment Region (IFN-SAR) on Expression of Vascular Endothelial Growth Factor-Fc (VEGF-Fc) Fusion Protein Expression in Chinese Hamster Ovary (CHO) Cells

“…IFN-SAR element is one of the most studied SARs, which have been successfully employed in both plasmid-based and virusbased vectors. 16,23 Our findings from stable VEGFR-Fc expression in cell pools and clones were in agreement with the results of previous studies, with 2-fold enhancement in cell-specific productivity. The same effect was observed in the clonal cells derived from cell pools.…”

The CAG promoter maintains high‐level transgene expression in HEK293 cells

“…The pEGFP-eGFP-matrix attachment region (MAR) plasmid, which contains the CMV promoter and MAR element, was previously constructed in our laboratory [31][32][33][34][35][36]. The CAG (GenBank accession no: GU299216.1, position 3-1664), HEF-1a (accession no: AY188393.1, position 10964-12623), CMV mutant, CHEF-1a (accession no: KY447299.1, position 12-1346), CAG enhancer (accession no: AJ575208.1, position 100-386), mouse CMV (accession no: KT343252.1, position 1103-1625), and PGK (accession no: KJ175229.1, position 641-1195) [37] promoters were synthesized and used to replace the CMV promoter in the pEGFP-eGFP vector (Fig.…”

Feasibility of chitosan-alginate (Chi-Alg) hydrogel used as scaffold for neural tissue engineering: a pilot studyin vitro

“…E-matrix is able to sustain cells and complex clusters of cells [37]. One advantage of E-matrix is its ability to immobilize water at appropriate storage temperatures and providing binding sites for cells, including nerve cells [38]. The increased level of polar amino-acid groups allows cells stored in E-matrix to be directly injected into a host organism without recognition by the host's immune system.…”