title: An Enterovirus-Like RNA Construct for Colon Cancer Suicide Gene Therapy.
abstract: In gene therapy, the use of RNA molecules as therapeutic agents has shown advantages over plasmid DNA, including higher levels of safety. However, transient nature of RNA has been a major obstacle in application of RNA in gene therapy.
  
  
  
  Here, we used the internal ribosomal entry site of encephalomyocarditis virus and the 3' non-translated region of Poliovirus to design an enterovirus-like RNA for the expression of a reporter gene (enhanced green fluorescent protein) and a suicide gene (thymidine kinase of herpes simplex virus). The expression of these genes was evaluated by flow cytometry and cytotoxicity assay in human colorectal adenocarcinoma cell line (SW480). We then armed RNA molecules with a target sequence for hsa-miR-143 to regulate their expression by microRNA (miRNA) mimics.
  
  
  
  The results showed effective expression of both genes by Entrovirus-like RNA constructs. The data also showed that the restoration of hsa-miR-143 expression in SW480 leads to a significant translation repression of the introduced reporter and suicide genes.
  
  
  
  Collectively, our data suggest the potential use of Entrovirus-like RNA molecules in suicide gene therapy. Additionally, as a consequence of the possible downregulated miRNA expression in cancerous tissues, a decreased expression of gene therapy constructs armed with target sequences for such miRNA in cancer tissue is expected.
authors.affiliation: Dept. of Virology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Rasekhian
authors.affiliation: Dept. of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Ladan
authors.initials: L
authors.lastname: Teimoori-Toolabi
authors.affiliation: Dept. of Hepatitis and HIV, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Safieh
authors.initials: S
authors.lastname: Amini
authors.affiliation: Dept. of Virology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Kayhan
authors.initials: K
authors.lastname: Azadmanesh
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000628:therapy*; D004770:Enterovirus / Q000235:genetics*; D015316:Genetic Therapy; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D050130:Oncolytic Virotherapy; D010957:Plasmids; D012367:RNA, Viral / Q000235:genetics*
chemical_list: D012367:RNA, Viral; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Polio 3’NCR; Thymidine kinase; miR-143