title: A new version of targeted minicircle producer system for EBV-positive human nasopharyngeal carcinoma.
abstract: Targeted gene therapy needs to be implemented for future therapies to ensure efficient activity at the site of patient primary tumors or metastases without causing intolerable side-effects. One of the elements of gene therapy is vector, which includes viral and non-viral vector. In the present study, we constructed a novel non-viral targeted gene therapeutic system by using the new minicircle (MC) producing plasmid for Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC). Molecular cloning technique was used to construct plasmids and electrophoretic analysis. Dual-luciferase reporter assay was used to evaluate the expression of luciferase. Fluorescence microscope was used to detect the expression of enhanced green fluorescence protein (EGFP). We constructed a new MC producing system pMC.BESPX-origin of plasmid replication (oriP), and demonstrated that this system could produce highly purified MC-oriP. Furthermore, our results showed that MC-oriP vector produced by the new system could mediate targeted luciferase gene expression in EBV-positive NPC cells. In addition, we verified that MC could mediate enhanced transgene expression compared with parent plasmid through EGFP transfection. The present study constructed a targeted expression vector pMC.BESPX-oriP which could carry diversified therapeutic genes for EBV-positive NPC and provides a new approach for MC-based therapies.
authors.affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, P.R. China.
authors.forename: Yufang
authors.initials: Y
authors.lastname: Zuo
authors.affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, P.R. China.
authors.forename: Sihai
authors.initials: S
authors.lastname: Liao
authors.affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, P.R. China.
authors.forename: Zumin
authors.initials: Z
authors.lastname: Xu
authors.affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, P.R. China.
authors.forename: Jierong
authors.initials: J
authors.lastname: Xie
authors.affiliation: State Key Laboratory of Oncology in South China, Cancer Center, Sun Yat-Sen University, Guangzhou, Guangdong, P.R. China.
authors.forename: Wenlin
authors.initials: W
authors.lastname: Huang
authors.affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, P.R. China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Yu
mesh_terms: D045744:Cell Line, Tumor; D004270:DNA, Circular / Q000235:genetics; D020031:Epstein-Barr Virus Infections / Q000150:complications / Q000628:therapy*; D017930:Genes, Reporter; D015316:Genetic Therapy; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D009303:Nasopharyngeal Neoplasms / Q000628:therapy* / Q000821:virology; D010957:Plasmids / Q000235:genetics; D014162:Transfection
chemical_list: D004270:DNA, Circular; D008156:Luciferases