ANTISENSE PHOSPHOROTHIOATE OLIGONUCLEOTIDES - SELECTIVE KILLING OF THE INTRACELLULAR PARASITE LEISHMANIA-AMAZONENSIS

被引:43
作者
RAMAZEILLES, C [1 ]
MISHRA, RK [1 ]
MOREAU, S [1 ]
PASCOLO, E [1 ]
TOULME, JJ [1 ]
机构
[1] UNIV BORDEAUX 2,INSERM,U386,MOLEC BIOPHYS LAB,F-33076 BORDEAUX,FRANCE
关键词
PROTOZOAN PARASITE; LOW DENSITY LIPOPROTEINS; HETERODUPLEX STABILITY; RNASE H;
D O I
10.1073/pnas.91.17.7859
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We targeted the mini-exon sequence, present at the 5' end of every mRNA of the protozoan parasite Leishmania amazonensis, by phosphorothioate oligonucleotides. A complementary 16-mer (16PS) was able to kill amastigotes-the intracellular stage of the parasite-in murine macrophages in culture. After 24 hr of incubation with 10 mu M 16PS, about 30% infected macrophages were cured. The oligomer 16PS acted through antisense hybridization in a sequence-dependent way; no effect on parasites was observed with noncomplementary phosphorothioate oligonucleotides. The antisense oligonucleotide 16PS was a selective killer of the protozoans without any detrimental effect to the host macrophage. Using 16PS linked to a palmitate chain, which enabled it to complex with low density lipoproteins, improved the leishmanicidal efficiency on intracellular amastigotes, probably due to increased endocytosis. Phosphorothioate oligonucleotides complementary to the intron part of the mini-exon pre-RNA were also effective, suggesting that antisense oligomers could prevent trans-splicing in these parasites.
引用
收藏
页码:7859 / 7863
页数:5
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