Ribozyme targeting of HIV-1 LTR.
The 5'-TAR region of HIV-1 mRNA is highly conserved amongst different HIV-1 isolates. We thus investigated the potential for in vivo targeting of the TAR RNA element by a hammerhead ribozyme. The use of the CAT reporter gene linked to the HIV1-LTR, in transient assays, reveals that a hammerhead ribozyme directed towards the first GUC of HIV-1 mRNA can efficiently inhibit CAT protein expression. We show that this inhibition is sequence-specific and probably due to a cleavage activity rather than an antisense effect. We show also that a hammerhead ribozyme that is inactive in vitro is capable of inhibiting CAT protein expression in a cellular environment. These results suggest that the targeting of the HIV-1 LTR by a hammerhead ribozyme constitutes a viable approach for anti-HIV therapy.
Ventura
M
M
ICGM, INSERM U 363, Université Paris V, Hospital Cochin, Paris, France.
Wang
P
P
Franck
N
N
Saragosti
S
S
D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D015870:Gene Expression; D015696:Gene Products, tat / Q000235:genetics / Q000494:pharmacology; D017930:Genes, Reporter; D016325:HIV Long Terminal Repeat / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D016337:RNA, Catalytic / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014162:Transfection; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D016337:RNA, Catalytic; D012333:RNA, Messenger; D054322:tat Gene Products, Human Immunodeficiency Virus; D015500:Chloramphenicol O-Acetyltransferase