RNA Interference in Mammalian Cells by Chemically-Modified RNA

Braasch, Dwaine A.; Jensen, Susan E.; Liu, Yinghui; Kaur, Kiran; Arar, Khalil; White, Michael A.; Corey, David R.

doi:10.1021/bi0343774

Cited by 503 publications

(461 citation statements)

References 40 publications

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“…Moreover, introduction of LNA into siRNA could result in significantly less off target-regulated genes (Elmen et al, 2005;Dahlgren et al, 2006;Mook et al, 2007). In agreement with other reports (Braasch et al, 2003;Elmen et al, 2005;Mook et al, 2007), we showed that our ERK2 LNA are substantially compatible with the siRNA machinery, exhibit improved biostability and enhance inhibition in vivo. Indeed, at least 15 days after transfection, phospho-ERK2 inhibition was observed with the ERK2 Stab1, in vivo.…”

Section: Erk2 Targeting Inhibits Tumor Growthsupporting

confidence: 92%

RNAi-mediated ERK2 knockdown inhibits growth of tumor cells in vitro and in vivo

et al. 2008

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The MAPK MEK/ERK pathway is often upregulated in cancer cells and represents an attractive target for development of anticancer drugs. Only few data concerning the specific functions of ERK1 and 2 are reported in the literature. In this report, we investigated the specific role of ERK1 and 2 in liver tumor growth both in vitro and in vivo. DNA synthesis and cells in S phase analysed by flow cytometry, correlated with strong inhibition of Cdk1 and cyclin E levels, are strongly reduced after exposure to the MEK inhibitor, U0126. We obtained a significant reduction of colony formation in soft agar assays and a reduction in the size of tumor xenografts in nude mice treated with U0126. Then, we could specifically abolished ERK1 or 2 expression by small-interfering RNA (siRNA) and demonstrated that ERK2 knockdown but not ERK1 interferes with the process of replication. Moreover, we found that colony formation and tumor growth in vivo were significantly inhibited by targeting ERK2 using stable chemically modified siRNA. Taken together, our results emphasize the importance of the MEK/ERK pathway in liver cancer cell growth in vitro and in vivo and argue for a crucial role of ERK2 in this regulation.

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Section: Erk2 Targeting Inhibits Tumor Growthsupporting

confidence: 92%

RNAi-mediated ERK2 knockdown inhibits growth of tumor cells in vitro and in vivo

et al. 2008

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“…In contrast, the adenovirus-associated viruses have been more effective in 92 Hydrodynamic injections of siRNAs alone or in conjunction with reporter constructs have led to the silencing of endogenous genes in various animal tissues, which include liver, spleen, lung, kidney and pancreas. [93][94][95][96] However, the need for a large volume that can be equivalent to up to 10-20% of the animals' blood volume limits this method of delivery in vivo. siRNAs, like most antisense oligomers, tend to be more bioavailable in the liver after in vivo administration.…”

Section: Rnai Deliverymentioning

confidence: 99%

siRNA-based approaches in cancer therapy

2006

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The availability of the human genome sequence has revolutionized the strategy of employing nucleic acids with sequences complementary to specific target genes to improve drug discovery and target validation. Development of sequence-specific DNA or RNA analogs that can block the activity of selected single-stranded genetic sequences offers the possibility of rational design with high specificity, lacking in many current drug treatments for various diseases including cancer, at relatively inexpensive costs. Antisense technology is one such example that has shown promising results and boasts of yielding the only approved drug to date in the genomics field. However, in vivo delivery issues have yet to be completely overcome for widespread clinical applications. In contrast to antisense oligonucleotides, the mechanism of silencing an endogenous gene by the introduction of a homologous double-stranded RNA (dsRNA), transgene or virus is called post-transcriptional gene silencing (PTGS) or RNA interference. PTGS is a natural mechanism whereby metazoan cells suppress expansion of genes when they come across dsRNA molecules with the same sequence. Short interfering RNA is currently the fastest growing sector of this antigene field for target validation and therapeutic applications. Although, in theory, the development of genomics-based agents to inhibit gene expression is simple and straightforward, the fundamental concern relies upon the capacity of the oligonucleotide to gain access to the target RNA. This paper summarizes the advances in the last decade in the field of PTGS using RNA interference approaches and provides relevant comparisons with other oligonucleotide-based approaches with a specific focus on oncology applications. Cancer Gene Therapy (2006) Genomic-based strategiesThe American Cancer Society estimates that, in 2005, a total of 1 372 910 new cancer cases and 570 280 deaths are expected in the United States. 1 These morbid statistics demonstrate the necessity of newer therapeutic modalities for achieving successful cancer treatment and cure. Downregulation of genes that contribute to cancer progression has been the goal of targeted genomics-based strategies, with the expectation that such an approach may lead to selective and specific inhibition of tumor growth with minimal untoward side effects on normal cells. Identification of target genes involved in neoplastic transformation and tumor progression has triggered the idea that nucleotide sequences of cancer-relevant genes could lead to the development of tailored anticancer agents that lack many of the toxic side effects of traditional cytotoxic drugs. This has led to a recent acceleration in the development and optimization of genomic-based strategies for cancer therapy. This idea dates back to the 1960s when RNA sequences were shown to serve as endogenous inhibitors of gene expression in procaryotes. The antigene approaches include the following:(1) Ribozymes: 2,3 These were discovered in the 1970s, and the elucidation of the transactivating hammerh...

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“…To enhance the stability of the chimeric RNAs in cell culture and in vivo 4,34,35,36,37 , the aptamer and sense strand segment of the siRNAs contained nuclease-resistant 2'-Fluoro UTP and 2'-Fluoro CTP and were synthesized from corresponding dsDNA templates by in vitro bacteriophage transcription (Fig 1). To prepare the siRNA containing chimeras, in vitro transcribed chimeric aptamer-sense strand polymers were annealed with equimolar concentrations of an unmodified antisense strand RNA.…”

Section: Design Of Anti-gp120 Aptamer-sirna Chimerasmentioning

confidence: 99%

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

Zhou

et al. 2007

Nat Prec

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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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RNA Interference in Mammalian Cells by Chemically-Modified RNA

Cited by 503 publications

References 40 publications

RNAi-mediated ERK2 knockdown inhibits growth of tumor cells in vitro and in vivo

RNAi-mediated ERK2 knockdown inhibits growth of tumor cells in vitro and in vivo

siRNA-based approaches in cancer therapy

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

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