Li Xiaojuan, Kuang Ersheng, Dai Wei, Zhou Boping, Yang Fuhua
Section of Molecular Virology, State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China.
Virus Res. 2005 Dec;114(1-2):126-32. doi: 10.1016/j.virusres.2005.06.005. Epub 2005 Jul 27.
Although it has been suggested that hepatitis delta virus (HDV) can be used as a vector to deliver biologically active RNAs into hepatocytes, modified HDV as a specific transporting and replicating vector in anti-viral research has not been investigated. In this study, we focused on the development of HDV as a replicative vector to deliver hammerhead ribozyme into hepatocytes and the study of the roles of delivered hammerhead ribozyme on the replication of hepatitis B virus (HBV). To investigate the effects of ribozyme delivered by HDV on HBV replication, we designed two hammerhead ribozymes that specifically target the hepatitis B virus genome. These two ribozymes were then inserted into the genome of hepatitis delta virus. Results showed that transfection of cells with tandem modified HDV cDNA resulted in the production of monomer form of sense and anti-sense genomic RNA indicating the recombinant HDV-ribozyme could replicate effectively. Our data also indicated that ribozymes delivered by the modified HDV had higher level of inhibition activity against HBV replication than that of ribozyme alone. This system provides a new approach for the study of mechanisms of HBV replication as well as for the potential treatment of HBV infection.
尽管有人提出丁型肝炎病毒（HDV）可作为载体将生物活性RNA导入肝细胞，但作为抗病毒研究中一种特异性转运和复制载体的修饰HDV尚未得到研究。在本研究中，我们专注于开发HDV作为复制载体将锤头状核酶导入肝细胞，并研究导入的锤头状核酶对乙型肝炎病毒（HBV）复制的作用。为了研究HDV递送的核酶对HBV复制的影响，我们设计了两种特异性靶向乙型肝炎病毒基因组的锤头状核酶。然后将这两种核酶插入丁型肝炎病毒基因组。结果表明，用串联修饰的HDV cDNA转染细胞导致产生有义链和反义链基因组RNA的单体形式，表明重组HDV-核酶能够有效复制。我们的数据还表明，修饰HDV递送的核酶对HBV复制的抑制活性水平高于单独的核酶。该系统为研究HBV复制机制以及潜在治疗HBV感染提供了一种新方法。
