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) |