Sarver N, Cantin E M, Chang P S, Zaia J A, Ladne P A, Stephens D A, Rossi J J
Division of Research and Development Program, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Science. 1990 Mar 9;247(4947):1222-5. doi: 10.1126/science.2107573.
Certain RNA molecules, called ribozymes, possess enzymatic, self-cleaving activity. The cleavage reaction is catalytic and no energy source is required. Ribozymes of the "hammerhead" motif were identified in plant RNA pathogens. These ribozymes possess unique secondary (and possibly tertiary) structures critical for their cleavage ability. The present study shows precise cleavage of human immunodeficiency virus type 1 (HIV-1) sequences in a cell-free system by hammerhead ribozymes. In addition to the cell-free studies, human cells stably expressing a hammerhead ribozyme targeted to HIV-1 gag transcripts have been constructed. When these cells were challenged with HIV-1, a substantial reduction in the level of HIV-1 gag RNA relative to that in nonribozyme-expressing cells, was observed. The reduction in gag RNA was reflected in a reduction in antigen p24 levels. These results suggest the feasibility of developing ribozymes as therapeutic agents against human pathogens such as HIV-1.
某些被称为核酶的RNA分子具有酶促自我切割活性。切割反应具有催化性，不需要能量来源。在植物RNA病原体中鉴定出了“锤头”基序的核酶。这些核酶具有对其切割能力至关重要的独特二级（可能还有三级）结构。本研究表明，在无细胞系统中，锤头核酶能精确切割1型人类免疫缺陷病毒（HIV-1）序列。除了无细胞研究外，还构建了稳定表达靶向HIV-1 gag转录本的锤头核酶的人类细胞。当用HIV-1攻击这些细胞时，相对于不表达核酶的细胞，观察到HIV-1 gag RNA水平大幅降低。gag RNA的减少反映在抗原p24水平的降低上。这些结果表明开发核酶作为针对HIV-1等人类病原体的治疗剂是可行的。
