DNA-enzyme-mediated cleavage of human immunodeficiency virus type 1 Gag RNA is significantly augmented by antisense-DNA molecules targeted to hybridize close to the cleavage site.
DNA-enzymes (Dzs) usually cleave short synthetic target RNAs very efficiently, but this activity diminishes significantly when tested on full-length RNAs, primarily because of the rigid secondary structures near the target sequence. We identified two Dzs, one each for 81-17 and 10-23 Dz, which cleaved the human immunodeficiency virus type 1 (HIV-1) Gag RNA poorly. We sought to use short oligodeoxynucleotides (ODNs) with the hope that it will facilitate Dz-mediated cleavage. The efficiencies of several ODNs were analyzed for their ability to augment the 8-17 Dz-mediated cleavage. We observed that ODNs that hybridized close to 5' and 3' ends of the target sequence were able to enhance significantly 8-17 Dz-mediated cleavage activity in a dose-dependent manner. The same was true for 10-23 Dz with ODNs that hybridized close to the target site. Thus, it was possible to enhance significantly the cleavage activity of poorly cleaving HIV-1 Gag-specific Dzs by using sequence-specific ODNs. This combination of antisense and catalytic Dz will, in principle, result in more effective gene suppression that could be exploited for therapeutic purposes.
Sood
Vikas
V
National Institute of Immunology, Department of Virology, New Delhi-110067, India.
Gupta
Nidhi
N
Bano
Aalia Sahar
AS
Banerjea
Akhil C
AC
D001483:Base Sequence; D016373:DNA, Antisense / Q000737:chemistry*; D021881:DNA, Catalytic / Q000737:chemistry / Q000378:metabolism*; D015750:Genes, gag; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism
D016373:DNA, Antisense; D021881:DNA, Catalytic; D020319:Oligodeoxyribonucleotides, Antisense; D012367:RNA, Viral; D016914:Ribonuclease H