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Alkylation of template strand of coding region causes effective gene silencing.

Abstract：

:We recently developed a new type of pyrrole (Py)-imidazole (Im) polyamide-tetrahydrocyclopropabenzindolone (CBI) conjugate with an indole linker as a stable sequence-specific alkylating agent. In this study, we investigated the gene silencing activities of polyamides A, B and C, which selectively alkylate specific sequences in the promoter region, non-coding strand and coding strand, respectively, of the green fluorescent protein (GFP) gene. GFP vectors were transfected into human colon carcinoma cells (HCT116), and the cells were treated with 100 nM of the polyamides for 24 h. Fluorescence microscopy indicated that a significant reduction of GFP fluorescence was only observed in the cells that were treated with polyamide C. In clear contrast, polyamides A and B did not show such activity. Moreover, real-time PCR demonstrated selective reduction of the expression of GFP mRNA following treatment with polyamide C. These results suggest that alkylating Py-Im polyamides that target the coding strand represent a novel approach for sequence-specific gene silencing.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Shinohara K,Sasaki S,Minoshima M,Bando T,Sugiyama H

doi

10.1093/nar/gkl005

keywords:

subject

Has Abstract

pub_date

2006-02-25 00:00:00

pages

1189-95

issue

4

eissn

0305-1048

issn

1362-4962

pii

34/4/1189

journal_volume

34

pub_type

杂志文章

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