ArticleScientific reports2018
Regulatory elements in vectors containing the ctEF-1α first intron and double enhancers for an efficient recombinant protein expression system.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 9 citations in OpenAlex.
- An effective method to enhance transient gene expression by transfecting high-density CHO cells.AMB Express · 2025Article
- Subnuclear organization and mislocalization of plasmids reduce transgene expression.Molecular therapy. Nucleic acids · 2025Article
- Optimization of the intron sequences combined with the CMV promoter increases recombinant protein expression in CHO cells.Scientific reports · 2025Article
- Construction of pVAX-1-based linear covalently closed vector with improved transgene expression.Molecular biology reports · 2024Article
- Article
- Enhanced Transgene Expression by Optimization of Poly A in Transfected CHO Cells.Frontiers in bioengineering and biotechnology · 2022Article
- Long noncoding RNA HAR1A regulates oral cancer progression through the alpha-kinase 1, bromodomain 7, and myosin IIA axis.Journal of molecular medicine (Berlin, Germany) · 2021Article
- The CAG promoter maintains high-level transgene expression in HEK293 cells.FEBS open bio · 2021Article
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Authors and funding
6 authors at 3 institutions in 2 countries.
Funding
Abstract
To establish a stable and scalable transient protein production system, we modified the EF-1 first intron size and verified the order of two recombinant enhancers downstream of the SV40 polyA sequence. This new vector was named pHH-Gemini (pHH-GM1) and was used to express alpha kinase 1 (ALPK1) and various other proteins, NLRP3, F-actin, Camodulin, PP2A, URAT1, Rab11a and myosin IIA. The results showed that, compared with six commercial plasmids, pHH-GM1 significantly enhanced His-HA-ALPK1 expression in a western blot analysis of transfected HEK293T cells. The expression of various other genes was also successful using the pHH-GM1 vector. In addition, we inserted turbo green florescence protein (tGFP) into the pHH-GM1 vector, and an improvement in fluorescence intensity was observed after transient transfection of HEK293T cells. For large-scale production, protein production was tested by standard supplementation with one volume of medium, and volumetric yields of 2 and 2.3 mg/L were achieved with pHH-GM1-ALPK1 in HEK293-F and CHO-S cells, respectively. We found that cell viability was more than 70% 11 days after cells were transfected with the pHH-GM1 vector. The pHH-GM1 vector with the ctEF-1α first intron and double enhancers, Simian virus 40 and Cytomegalovirus (SV40 and CMV) is an efficient CMV promoter-based gene expression system that can potentially be applied to study genes of interest and improve protein production.
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