Phosphorothiolate ribozyme against the conserved sequence in V3 loop of HIV-1.
Fluorescent-labeled phosphorothiolate DNA-RNA chimeric hammerhead ribozyme (thio-Rz) was synthesized. The ribozyme recognizes relatively conserved sequences in the functionally important region (V3 loop) of envelope protein of HIV-1. The thio-Rz as well as wild-type ribozyme cleaved RNA transcripts (414 base) of V3 region in a dose-dependent manner. After delivery into the cells, both wild-type and thio-Rz were able to be readily detected. The positive signals faded within 12 h in the former case, while they lasted 24 h in the latter case. The inhibitory effect by thio-Rz on cell fusion between envelope-expressing CHO cells and CD4+ cells appeared to be time-dependent, and about 70% of inhibition was seen at 72 h. Possible applications of the ribozyme against HIV-1 infection were discussed.
Zhang
X
X
Laboratory of AIDS Immunology, Kyoto University, Japan.
Yamada
R
R
Shimayama
T
T
Imada
K
K
Uchiyama
T
T
Hattori
T
T
D001483:Base Sequence; D017124:Conserved Sequence; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009946:Organothiophosphorus Compounds; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*
D015699:HIV Envelope Protein gp120; D009946:Organothiophosphorus Compounds; D016337:RNA, Catalytic