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• W1998349008 endingPage "476" @default.
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• W1998349008 abstract "Ribonuclease H (RNase H), an enzyme that cleaves an RNA sequence base-paired with a complementary DNA sequence, is proposed to be the mediator of antisense phosphorothioate oligonucleotide (S-oligo) lethality in a cell. To understand the role of RNase H in the killing of the parasitic protozoan Leishmania by antisense S-oligos, we expressed an episomal copy of the Trypanosoma brucei RNase H1 gene inside L. amazonensis promastigotes and amastigotes that constitutively express firefly luciferase. Our hypothesis was that S-oligo-directed degradation of target mRNA is facilitated in a cell that has higher RNase H activity. Increased inhibition of luciferase mRNA expression by anti-luciferase S-oligo and by anti-miniexon S-oligo in these stably transfected promastigotes overexpressing RNase H1 was correlated to the higher activity of RNase H in these cells. The efficiency of killing of the RNase H overexpressing amastigotes inside L. amazonensis-infected macrophages by anti-miniexon S-oligo was higher than in the control cells. Thus, RNase H appears to play an important role in the antisense S-oligo-mediated killing of Leishmania. Chemical modification of S-oligos that stimulate RNase H and/or co-treatment of cells with an activator of RNase H may be useful for developing an antisense approach against leishmaniasis. The transgenic Leishmania cells overexpressing RNase H should be a good model system for the antisense-mediated gene expression ablation studies in these parasites." @default.
• W1998349008 created "2016-06-24" @default.
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• W1998349008 creator A5075612382 @default.
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• W1998349008 date "2001-02-01" @default.
• W1998349008 modified "2023-09-28" @default.
• W1998349008 title "Increased efficacy of antileishmanial antisense phosphorothioate oligonucleotides in Leishmania amazonensis overexpressing ribonuclease H11Abbreviations: RNase H, ribonuclease H; S-oligo, phosphorothioate-modified oligodeoxyribonucleotide; LUAS, anti-luciferase antisense S-oligo; LUS, S-oligo complementary to LUAS; ASM, S-oligo antisense to Leishmania miniexon; SSM, S-oligo complementary to ASM; HIFBS, heat-inactivated fetal bovine serum; TbRH1, Trypanosoma brucei RNase H1 gene; RACE, rapid …" @default.
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• W1998349008 doi "https://doi.org/10.1016/s0006-2952(00)00568-2" @default.
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