Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells

被引：310

作者：

Chiu, YL ^{[1
]}

Ali, A ^{[1
]}

Chu, CY ^{[1
]}

Cao, H ^{[1
]}

Rana, TM ^{[1
]}

机构：

[1] Univ Massachusetts, Sch Med, Dept Biochem & Mol Pharmacol, Worcester, MA 01605 USA

来源：

CHEMISTRY & BIOLOGY | 2004年 / 11卷 / 08期

关键词：

D O I：

10.1016/j.chembiol.2004.06.006

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

RNA interference (RNAi) is the process by which short-interfering RNA (siRNA) target a specific mRNA for degradation through interactions with an RNA-induced silencing complex (RISC). Here, a clear correlation between siRNA localization, cellular uptake, and RNAi activity was discovered by delivering siRNA into cells using siRNA-TAT(47-57) peptide, siRNA-TAT(47-57)-derived oligocarbamate conjugates, or nanoparticles. For successful RNAi, the localization of siRNA was distinctly perinuclear, suggesting that siRNA is targeted to these regions for interactions with RISC to induce RNAi. siRNA sequence variation and the presence of the target mRNA apparently did not change the subcellular localization pattern of siRNA. Intriguingly, siRNA conjugated to TAT(47-57) peptide or TAT(47-57)-derived oligocarbamate resulted in efficient RNAi activity and perinuclear localization of siRNA that was distinctly different from nonconjugated free TAT peptide nucleolar localization. These results suggest that interactions with RISC dictate siRNA localization even when siRNA is conjugated to TAT(47-57) peptide.

引用

页码：1165 / 1175

页数：11

共 52 条

[21] Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells [J].

Lee, NS ;

Dohjima, T ;

Bauer, G ;

Li, HT ;

Li, MJ ;

Ehsani, A ;

Salvaterra, P ;

Rossi, J .

NATURE BIOTECHNOLOGY, 2002, 20 (05) :500-505

[22] Cell-penetrating peptides [J].

Lindgren, M ;

Hallbrink, M ;

Prochiantz, A ;

Langel, U .

TRENDS IN PHARMACOLOGICAL SCIENCES, 2000, 21 (03) :99-103

[23] Synthetic DNA delivery systems [J].

Luo, D ;

Saltzman, WM .

NATURE BIOTECHNOLOGY, 2000, 18 (01) :33-37

[24] ENDOCYTOSIS AND TARGETING OF EXOGENOUS HIV-1 TAT PROTEIN [J].

MANN, DA ;

FRANKEL, AD .

EMBO JOURNAL, 1991, 10 (07) :1733-1739

[25] Gene silencing in mammals by small interfering RNAs [J].

McManus, MT ;

Sharp, PA .

NATURE REVIEWS GENETICS, 2002, 3 (10) :737-747

[26] U6 promoter-driven siRNAs with four uridine 3′ overhangs efficiently suppress targeted gene expression in mammalian cells [J].

Miyagishi, M ;

Taira, K .

NATURE BIOTECHNOLOGY, 2002, 20 (05) :497-500

[27] Translocating peptides and proteins and their use for gene delivery [J].

Morris, MC ;

Chaloin, L ;

Heitz, F ;

Divita, G .

CURRENT OPINION IN BIOTECHNOLOGY, 2000, 11 (05) :461-466

[28] HIV Tat peptide enhances cellular delivery of antisense morpholino oligomers [J].

Moulton, HM ;

Hase, MC ;

Smith, KM ;

Iversen, PL .

ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT, 2003, 13 (01) :31-43

[29] Transduction of full-length TAT fusion proteins into mammalian cells:: TAT-p27Kip1 induces cell migration [J].

Nagahara, H ;

Vocero-Akbani, AM ;

Snyder, EL ;

Ho, A ;

Latham, DG ;

Lissy, NA ;

Becker-Hapak, M ;

Ezhevsky, SA ;

Dowdy, SF .

NATURE MEDICINE, 1998, 4 (12) :1449-1452

[30] Basic peptides as functional components of non-viral gene transfer vehicles [J].

Nakanishi, M ;

Eguchi, A ;

Akuta, T ;

Nagoshi, E ;

Fujita, S ;

Okabe, J ;

Senda, T ;

Hasegawa, M .

CURRENT PROTEIN & PEPTIDE SCIENCE, 2003, 4 (02) :141-150

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