Fusion with matrix attachment regions enhances expression of recombinant protein in human HT-1080 cells.
J Biosci Bioeng
; 130(5): 533-538, 2020 Nov.
Article
in En
| MEDLINE
| ID: mdl-32773266
ABSTRACT
Like endogenous proteins, recombinant foreign proteins produced in human cell lines also need post-translational modifications. However, high and long-term expression of a gene of interest (GOI) presents significant challenges for recombinant protein production in human cells. In this work, the effect of human matrix attachment region elements (MARs), including the ß-globin MAR (gMAR), chicken lysozyme MAR (cMAR), and a combination of these two, on the stable expression of GOI was assessed in human HT-1080 cells. After transfection with vectors containing the MAR elements and eGFP, stably HT-1080 cell pools were obtained under selective pressure. eGFP protein expression was analyzed by flow cytometry, while transgene copy number and eGFP mRNA expression levels were determined with qPCR and qRT-PCR technology. We found that MARs could not enhance transfection efficiency, but gMAR could significantly increase eGFP expression in stable HT-1080 cell pools by approximately 2.69-fold. Moreover, gMAR could also increase eGFP expression stability during long-term culture. Lastly, we showed that the effect of the MARs on transgenes was related to the gene copy number. In summary, this study found that MARs could both enhance the transgene expression and stability in HT-1080 cells.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Recombinant Fusion Proteins
/
Genetic Engineering
/
Matrix Attachment Regions
Limits:
Humans
Language:
En
Journal:
J Biosci Bioeng
Journal subject:
ENGENHARIA BIOMEDICA
/
MICROBIOLOGIA
Year:
2020
Document type:
Article
Affiliation country:
China