Antiviral Evaluation of HIV-1 Specific Ribozyme Expressed in CD4+ HeLa Cells
Catalytic RNA, called ribozyme, has been shown to enzymatically cleave specific sites in RNA (for reviews, see Cech and Bass, 1986, and Uhlmann and Peymen, 1990). One group of ribozymes has an active site of consensus sequences forming a “hammerhead” structure. The active center of the hammerhead ribozyme consists of 11 essential bases juxtaposed near 3 target bases (Cech, 1988). The potential versatility of this system lies in the fact that simple ribozymes can be constructed which target sites in native RNA, completing the active site in a trans configuration, and inducing cleavage (Haseloff and Gerlach, 1988). Ribozymes, which have been developed to cleave human immunodeficiency virus type 1 (HIV-1) RNA, can be transcribed in both in vitro and in vivo systems (Chang et al., 1990; Sarver et al.,1990). The purpose of this report is to review the studies evaluating the effect of ribozyme on HIV-1 RNA cleavage and on the antiviral effect in mammalian cells. Using a hammerhead motif, ribozymes were constructed which targeted a gag cleavage site, were cloned into a mammalian expression vector, and used to transform HeLa-CD4+ cells. These cells expressed HIV-1-specific ribozyme and demonstrated inhibition of HIV-1 infection.
KeywordsDulbecco Modify Eagle Medium Hammerhead Ribozyme States Public Health Service Grant Ribozyme Expression Synthetic Ribozyme
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