title: Human miR-5193 Triggers Gene Silencing in Multiple Genotypes of Hepatitis B Virus.
abstract: Hepatitis B virus (HBV) infection can lead to various disease states including asymptomatic, acute hepatitis, chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC), which remain a major health problem worldwide. Previous studies demonstrated that microRNA (miRNAs) plays an important role in viral replication. This study aimed to predict and evaluate human miRNAs targeting multiple genotypes of HBV. Candidate human miRNAs were analyzed by data obtained from miRBase and RNAhybrid. Then miRNAs were selected based on hybridization patterns and minimum free energy (MFE). The silencing effect of miRNA was evaluated by real-time PCR, the luciferase reporter assay and the ELISA assay. Five human miRNAs including miR- 142-5p, miR-384, miR-500b, miR-4731-5p and miR-5193 were found to target several HBV genotypes. Interestingly, miR-5193 was found to be the most potent miRNA that could target against all HBV transcripts in almost all HBV genotypes with a highly stable hybridization pattern (5' canonical with MFE lower than -35 kcal/mol). Moreover, miR-5193 caused significant silencing in luciferase activity (53% reduction), luciferase transcript (60% reduction) and HBV surface antigen (HBsAg) production (20-40% reduction depending on genotypes). Therefore, miR-5193 might be useful and have a vital role for inhibition of HBV replication in the future.
authors.forename: Apichaya
authors.initials: A
authors.lastname: Khlaiphuengsin
authors.forename: Nattanan Panjaworayan
authors.initials: NP
authors.lastname: T-Thienprasert
authors.forename: Pisit
authors.initials: P
authors.lastname: Tangkijvanich
authors.forename: Nawarat
authors.initials: N
authors.lastname: Posuwan
authors.forename: Jarika
authors.initials: J
authors.lastname: Makkoch
authors.forename: Yong
authors.initials: Y
authors.lastname: Poovorawan
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Rama 4 Rd. Pathumwan, Bangkok, Thailand 10330. sp.medbiochemcu@gmail.com.
authors.forename: Sunchai
authors.initials: S
authors.lastname: Payungporn
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D015870:Gene Expression; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D005838:Genotype; D056945:Hep G2 Cells; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D034622:RNA Interference; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics
chemical_list: D006514:Hepatitis B Surface Antigens; C000609353:MIRN5193 microRNA, human; D035683:MicroRNAs; D012367:RNA, Viral