Ribozymes as potential anti-HIV-1 therapeutic agents.
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.
Sarver
N
N
Division of Research and Development Program, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Cantin
E M
EM
Chang
P S
PS
Zaia
J A
JA
Ladne
P A
PA
Stephens
D A
DA
Rossi
J J
JJ
D000163:Acquired Immunodeficiency Syndrome / Q000188:drug therapy*; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D015870:Gene Expression; D015683:Gene Products, gag / Q000378:metabolism; D015750:Genes, gag / Q000187:drug effects*; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D016337:RNA, Catalytic; D012335:RNA, Ribosomal / Q000494:pharmacology* / Q000627:therapeutic use; D012367:RNA, Viral / Q000187:drug effects*; D014162:Transfection; D014758:Viral Core Proteins / Q000378:metabolism
D015683:Gene Products, gag; D016655:HIV Core Protein p24; D016337:RNA, Catalytic; D012335:RNA, Ribosomal; D012367:RNA, Viral; D014758:Viral Core Proteins