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 β-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.

像内源蛋白一样，在人类细胞系中产生的重组外源蛋白也需要翻译后修饰。然而，目的基因（GOI）的长期和长期表达对人类细胞中重组蛋白的生产提出了重大挑战。在这项工作中，评估了人类基质附着区元素（MARs），包括β-珠蛋白MAR（gMAR），鸡溶菌酶MAR（cMAR）以及这两者的组合，对人GOI稳定表达的影响。 HT-1080细胞。用含有MAR元素和eGFP的载体转染后，在选择性压力下获得稳定的HT-1080细胞池。通过流式细胞仪分析eGFP蛋白的表达，而转基因拷贝数和eGFP用qPCR和qRT-PCR技术测定mRNA表达水平。我们发现，MARs不能提高转染效率，但gMAR可以显着增加稳定的HT-1080细胞池中的eGFP表达约2.69倍。此外，gMAR还可以在长期培养过程中提高eGFP表达的稳定性。最后，我们证明了MARs对转基因的影响与基因拷贝数有关。总而言之，这项研究发现MARs可以增强HT-1080细胞的转基因表达和稳定性。