ArticleMedical science monitor : international medical journal of experimental and clinical research2017
Using a Commonly Down-Regulated Cytomegalovirus (CMV) Promoter for High-Level Expression of Ectopic Gene in a Human B Lymphoma Cell Line.
Article in Medical science monitor : international medical journal of experimental and clinical research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
3 citing papers in PubMed, 7 citations in OpenAlex.
- Biopolymer-based Carriers for DNA Vaccine Design.Angewandte Chemie (International ed. in English) · 2021Review
- Searching for Promoters to Drive Stable and Long-Term Transgene Expression in Fibroblasts for Syngeneic Mouse Tumor Models.International journal of molecular sciences · 2020Article
- Retracted Article: Long noncoding RNA ANRIL protects cardiomyocytes against hypoxia/reoxygenation injury by sponging miR-195-5p and upregulating Bcl-2.RSC advances · 2019Article
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Authors and funding
7 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
BACKGROUND Vectors are widely used to drive gene expression using a promoter. However, not all promoters are able to drive ectopic gene expression efficiently, including CMV promoter. Here, we report our data using CMV promoter for high-level gene expression in a B lymphoma cell line DG75. MATERIAL AND METHODS A plasmid (pcDNA3.1(+)) containing the CD21 gene driven under CMV promoter was constructed. The plasmid was stably transfected into a human B lymphoma cell line DG75 for cellular surface CD21 expression, and flow cytometry was used to monitor CD21 expression. CD21+ cells in the stable cell line were purified using anti-CD21 antibody-coupled Dynabeads for CD21-mediated antigen presentation experiment. RESULTS The percentage of CD21+ cells in newly generated stable DG75-pcDNA3.1(+)-CD21 cells was only 6.5% as determined by flow cytometry, which was unexpected and did not fit the requirements for further experiments. However, CD21+ cells could be purified to 100% using anti-CD21 antibody-coupled beads. The percentage of CD21+ cells in purified cells can be kept at 95%, 82%, 42%, 15%, and 42% at 7 d, 14 d, 34 d, and 42 d after purification, respectively. Specific T cell response against CD21-mediated antigen presentation can be activated successfully only when surface CD21 expression remains high. CONCLUSIONS A commonly down-regulated CMV promoter can be used to drive ectopic gene expression at a high-level in stable cell lines. Our results should facilitate future experimental design using other down-regulated promoters containing vectors such as SV40 and PGK1.
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