Sequence‐specific gene silencing in murine muscle induced by electroporation‐mediated transfer of short interfering RNA

Kishida, Tsunao; Asada, Hidetsugu; Gojo, Satoshi; Ohashi, Suzuyo; Shin‐Ya, Masaharu; Yasutomi, Kakei; Terauchi, Ryu; Takahashi, Kenji; Kubo, Toshikazu; Imanishi, Jirô; Mazda, Osam

doi:10.1002/jgm.456

Cited by 72 publications

(55 citation statements)

References 47 publications

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“…Nonviral transfection with synthetic siRNA is free from virusassociated adverse effects, while the RNAi effect may be transient. 43,53,54 Although the most appropriate delivery system should be selected for each RNAi therapeutic application, electrotransfer of synthetic siRNA may be suitable for Mitf-knockdown therapy for melanoma, because long-lasting silencing effects are not always required to kill tumor cells that have been successfully transfected with siRNA.…”

Section: Discussionmentioning

confidence: 99%

“…55,56,[59][60][61] Electroporation was applied to RNAi by Pekarik et al 62 who transfected synthetic siRNA in ovo into chick embryos, while we first documented electrotransfer of siRNA into a mammalian organ in vivo. 53 We also employed electroporation to therapeutic RNAi for rheumatoid arthritis in rat, indicating that the articular disorder was drastically ameliorated by silencing tumor necrosis factor-a in inflammatory sinovial tissue. 43 An advantage of electroporation is that the siRNA can be exclusively introduced into the cells that reside in the portion between the electrodes.…”

Section: Discussionmentioning

confidence: 99%

“…After incubation for 5 h at 371C, the optical density (OD) at 570 nm was measured. Protein concentration in each extract was measured using residual 10 ml aliquots as described, 53 and the tyrosinase activity was standardized by the protein content.…”

Section: Tyrosinase Activitymentioning

confidence: 99%

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Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

et al. 2006

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Microphthalmia-associated transcription factor (Mitf) is critically involved in melanin synthesis as well as differentiation of cells of the melanocytic lineage. Some earlier studies suggested that Mitf is also essential in the survival of melanoma cells, but this notion remains controversial. We synthesized short interfering RNA (siRNA) duplexes corresponding to the mitf sequence and transfected them into B16 melanoma. Lipid-mediated transfection in vitro of Mitfspecific siRNA resulted in specific downregulation of Mitf and of the tyrosinase that is a transcriptional target of Mitf. This treatment also remarkably reduced the viability of melanoma cells by inducing apoptosis. To examine the potential feasibility of RNAi therapy against melanoma, B16 cells were subcutaneously injected into syngenic mice and siRNA was transfected into the pre-established tumor by means of electroporation. The Mitf-specific siRNA drastically reduced outgrowth of subcutaneous melanoma, while nonspecific siRNA failed to affect tumor progression. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-based analysis of tumor specimens demonstrated that the tumor cells transfected with Mitf-siRNA effectively underwent apoptosis in vivo. The present results indicate that Mitf plays important roles in melanoma survival. Intratumor electrotransfer of Mitf-specific siRNA may provide a powerful strategy for therapeutic intervention of malignant melanoma.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

et al. 2006

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“…In mammalian cells, short interfering RNA (siRNA) 21-23 nucleotide pairs in length silences the gene with the homologous sequence [23,24]. The in vivo effectiveness of siRNA-mediated silencing was also studied in various organs, including the liver [25,26] and the skeletal muscle [27]. However, in vivo RNAi in cardiac tissues has not been reported so far.…”

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confidence: 99%

Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart

Tsunoda

Mazda

Oda

et al. 2005

Biochemical and Biophysical Research Communications

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129

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Naked plasmid DNA (pDNA) and short interfering RNA (siRNA) duplexes were transduced into adult murine heart by means of sonoporation using the third-generation microbubble, BR14. Plasmid DNAs carrying luciferase, b-galactosidase (b-gal), or enhanced green fluorescent protein (EGFP) reporter genes were mixed with BR14 and injected percutaneously into the left ventricular (LV) cavity of C57BL/6 mice while exposed to transthoracic ultrasound at 1 MHz for 60 s. Sonoporation at an output intensity of 2.0 W/cm 2 and a 50% pulse duty ratio resulted in the highest luciferase expression in the heart. Histological examinations revealed significant expression of the b-gal and EGFP reporters in the subendocardial myocardium of LV. Intraventricular co-injection of siRNA-GFP and BR14 with concomitant ultrasonic exposure resulted in substantial reduction in EGFP expression in the coronary artery in EGFP transgenic mice. The present method may be applicable to gain-of-function and loss-of-function genetic engineering in vivo of adult murine heart.

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“…Currently, there are several methods to deliver siRNA in vivo. These can be divided into physical and mechanical methods (hydrodynamic tail vein injections in mice 4,5,6 , electroporation 7,8,9 , ultrasound 10 , and the gene gun 11 ); local administration 3 (intravenous injection 12 , intraperitoneal injection, subcutaneous injection); and chemical methods (cationic lipids 13,14 , polymers 15,16,17,18,19,20 , and peptides 21,22,23,24 ). However, the delivery efficiency (desired dose), uncontrollable biodistribution and delivery-related toxicitities must be carefully analyzed.…”

Section: Discussionmentioning

confidence: 99%

Novel Cell type-specific aptamer-siRNA delivery system for HIV-1 therapy

Zhou

et al. 2007

Nature Precedings

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SummaryThe successful use of small interfering RNAs (siRNAs) for therapeutic purposes requires safe and efficient delivery to specific cells and tissues. Here we demonstrate cell type-specific delivery of anti-HIV siRNAs via fusion to an anti-gp120 aptamer. The envelope glycoprotein is expressed on the surface of HIV-1 infected cells, allowing binding and interalization of the aptamer-siRNA chimeric molecules. We demonstrate that the anti-gp120 aptamer-siRNA chimera is specifically taken up by cells expressing HIV-1 gp120, and the appended siRNA is processed by Dicer, releasing an anti-tat/rev siRNA which in turn inhibits HIV replication. We show for the first time a dual functioning aptamer-siRNA chimera in which both the aptamer and the siRNA portions have potent anti-HIV activities and that gp120 expressed on the surface of HIV infected cells can be used for aptamer mediated delivery of anti-HIV siRNAs.RNA interference (RNAi) is a process of sequence-specific post-transcriptional gene silencing triggered by small interfering RNAs (siRNA). The silencing is sequence specific and one of the two strands of the siRNA guides the RNA induced silencing complex (RISC) to the complementary target, resulting in cleavage and subsequent destruction of the target RNA 1 . RNAi is rapidly becoming one of the methods of choice for gene function studies, and is also being exploited for therapeutic applications 2, 3 . The

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Sequence‐specific gene silencing in murine muscle induced by electroporation‐mediated transfer of short interfering RNA

Abstract: The present system may provide a useful means of phenotypic analysis of genetic information in mammalian organs for basic research as well as therapeutic molecular targeting in the post-genomic era.

Cited by 72 publications

References 47 publications

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

Therapeutic RNA interference of malignant melanoma by electrotransfer of small interfering RNA targeting Mitf

Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart

Novel Cell type-specific aptamer-siRNA delivery system for HIV-1 therapy

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