RRC ID 10734
著者 Matsui K, Horiuchi S, Sando S, Sera T, Aoyama Y.
タイトル RNAi silencing of exogenous and endogenous reporter genes using a macrocyclic octaamine as a "compact" siRNA carrier. Studies on the nonsilenced residual activity.
ジャーナル Bioconjug Chem
Abstract A macrocyclic octaamine 1 having a covalently linked lipid-bundle structure was introduced as a new type of siRNA carrier. Gel electrophoresis, DLS, and SPR results indicate that it strongly binds to a luciferase-targeting 21-mer (42P) siRNA with a ratio of 1/P congruent with 0.3 (1/N congruent with 2.4) to give remarkably compact 1-siRNA complexes with an average size of approximately 10 nm. The 1-mediated siRNA silencing of the exogenous luciferase gene occurs with a 90-95% efficiency. The overall suppression-[siRNA] profile with a 5-10% residual activity in the saturation region is commonly observed irrespective of the cell type (HeLa, HepG2, or HEK293), the order, or timing (stepwise or simultaneous) of supply of the siRNA and that of the luciferase-encoding plasmid, the level of mRNA transcribed, or the type of carriers (1 vs lipofectamine 2000). The silencing of the endogenous DsRed2 gene stably incorporated in the genome of HeLa cells also has a similar overall profile. These results suggest that (1) the cellular uptake of the plasmid and that of the siRNA are basically independent of each other and (2) the incomplete silencing is not due to insufficient siRNA delivery. Implication of item 2 is briefly discussed.
巻・号 17(1)
ページ 132-8
公開日 2006-1-1
DOI 10.1021/bc050112l
PMID 16417261
MeSH Cell Line DNA / metabolism Drug Carriers / chemistry Drug Carriers / metabolism* Genes, Reporter HeLa Cells Humans Macrocyclic Compounds / chemistry Macrocyclic Compounds / metabolism* Plasmids / genetics Plasmids / metabolism Polyamines / chemistry Polyamines / metabolism* RNA Interference* RNA, Small Interfering / chemistry RNA, Small Interfering / metabolism* Transfection
IF 4.031
引用数 13
WOS 分野 CHEMISTRY, MULTIDISCIPLINARY CHEMISTRY, ORGANIC BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 HeLa(RCB0007) 293(RCB1637)