Passman M, Weinberg M, Kew M, Arbuthnot P
Department of Molecular Medicine and Haematology, University of the Witwatersrand Medical School, 7 York Road, Parktown, 2193, South Africa.
Biochem Biophys Res Commun. 2000 Feb 24;268(3):728-33. doi: 10.1006/bbrc.2000.2209.
Chronic hepatitis B virus (HBV) infection is endemic to several populous areas of the world and is frequently complicated by hepatocellular carcinoma. Ribozymes can be designed to cleave target RNA sequences specifically and show promise for the treatment of HBV infection. Demonstration of intracellular inhibition of HBV gene expression, essential to developing therapeutic ribozymes, has been the aim of this investigation. We generated two vectors encoding hammerhead ribozymes that target the HBx region of HBV. Plasmids containing intact HBV sequences or a modification in which the preS2/S region was replaced by DNA encoding enhanced green fluorescent protein (EGFP) were used to test ribozyme action in transfected cells. Both ribozymes inhibited surface antigen secretion and EGFP expression similarly. The measurement of EGFP expression is convenient to assess ribozyme action in situ and effective targeting of HBV sequences that are common to all HBV transcripts is potentially useful to develop strategies to counter HBV infection.
慢性乙型肝炎病毒（HBV）感染在世界上几个人口众多的地区呈地方性流行，并且常常并发肝细胞癌。核酶可以被设计成特异性切割靶RNA序列，显示出治疗HBV感染的前景。证明对HBV基因表达的细胞内抑制作用，这对开发治疗性核酶至关重要，一直是本研究的目标。我们构建了两个编码靶向HBV的X区的锤头状核酶的载体。含有完整HBV序列的质粒或其中前S2/S区被编码增强型绿色荧光蛋白（EGFP）的DNA取代的修饰质粒，用于测试转染细胞中的核酶作用。两种核酶对表面抗原分泌和EGFP表达的抑制作用相似。EGFP表达的测量便于原位评估核酶作用，有效靶向所有HBV转录本共有的HBV序列可能有助于制定对抗HBV感染的策略。
