Inhibition of Plasmodium falciparum malaria using antisense oligodeoxynucleotides.
We studied inhibition of growth of the malaria parasite Plasmodium falciparum in in vitro culture using antisense (AS) oligodeoxynucleotides (ODNs) against different target genes. W2 and W2mef strains of drug-resistant parasites were exposed to AS ODNs over 48 hr, and growth was determined by microscopic examination and [3H]hypoxanthine incorporation. At ODN concentrations of 1 microM, phosphorothioate (PS) ODNs inhibited growth in a target-independent manner. However, between 0.5 and 0.005 microM, ODNs against dihydrofolate reductase, dihydropteroate synthetase, ribonucleotide reductase, the schizont multigene family, and erythrocyte binding antigen EBA175 significantly inhibited growth compared with a PS AS ODN against human immunodeficiency virus, two AS ODNs containing eight mismatches, or the sense strand controls (P < 0.0001). The IC50 was approximately 0.05 microM, whereas that for non-sequence-specific controls was 15-fold higher. PS AS ODNs against DNA polymerase alpha showed less activity than that for other targets, whereas a single AS ODN against triose-phosphate isomerase did not differ significantly from controls. We conclude that at concentrations below 0.5 microM, PS AS ODNs targeted against several malarial genes significantly inhibit growth of drug-resistant parasites in a nucleotide sequence-dependent manner. This technology represents an alternative method for identifying malarial genes as potential drug targets.
Barker
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Hybridon, Inc., Worcester, MA 01605, USA.
Metelev
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Rapaport
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Zamecnik
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D000818:Animals; D001483:Base Sequence; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D017125:Genes, Protozoan; D016778:Malaria, Falciparum / Q000628:therapy; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010963:Plasmodium falciparum / Q000201:enzymology / Q000254:growth & development*; D015800:Protozoan Proteins / Q000235:genetics; D012264:Ribonucleotide Reductases / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014305:Triose-Phosphate Isomerase / Q000235:genetics
D016376:Oligonucleotides, Antisense; D015800:Protozoan Proteins; D012264:Ribonucleotide Reductases; D013762:Tetrahydrofolate Dehydrogenase; D004259:DNA-Directed DNA Polymerase; D014305:Triose-Phosphate Isomerase