title: An aptamer targets HBV core protein and suppresses HBV replication in HepG2.2.15 cells.
abstract: Hepatitis B virus (HBV)-related hepatitis is a major health concern worldwide. As current anti-HBV therapies are limited, it is essential to develop new strategies. Aptamer, a newly developed adaptive molecule (single-strand DNA or RNA also known as nucleotide antibody), is a new strategy for clinical diagnosis and therapy due to its high affinity and specificity. In the present study, by systematic evolution of ligand by exponential enrichment (SELEX), aptamers were screened against the core protein of HBV (HBc) from a random ssDNA library. Quantitative PCR (qPCR) results showed that the binding proportions of the SELEX-enriched aptamer pools were increased with the SELEX rounds, until round seven. Thus, the pool of round seven was cloned. Following the sequence analysis of a total of 90 clones by Macaw software, five aptamer candidates were selected and their affinity to HBc was tested by dot blot. Apt.No.28, which had sequence replicates in the clones, was shown to have a high affinity. Furthermore, by agarose gel electrophoresis-capillary transfer-blotting and qPCR, Apt.No.28 was shown to inhibit the assembly of the nucleocapsid, reducing extracellular HBV DNA whose synthesis relied on the formation of the nucleocapsid, indicating its role in suppressing HBV replication. The results provided a new ideal targeting molecule and may facilitate the strategy for targeted therapy as well as drug development of HBV-related diseases.
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Zuowei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Pei
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Lu
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Xiaojiao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, P.R. China.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
mesh_terms: D052157:Aptamers, Nucleotide / Q000494:pharmacology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000235:genetics*; D004279:DNA, Viral / Q000302:isolation & purification; D015723:Gene Library; D056945:Hep G2 Cells; D006512:Hepatitis B Core Antigens / Q000737:chemistry*; D006514:Hepatitis B Surface Antigens / Q000302:isolation & purification; D006513:Hepatitis B e Antigens / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D016415:Sequence Alignment; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens