Small interfering RNA treatment can inhibit Cyprinid herpesvirus 3 associated cell death in vitro

2014 ◽  
Vol 17 (4) ◽  
pp. 733-735 ◽  
Author(s):  
M. Adamek ◽  
G. Rauch ◽  
G. Brogden ◽  
D. Steinhagen
Keyword(s):  
Cell Death ◽  
Rna Interference ◽  
Small Interfering Rna ◽  
Enzyme Synthesis ◽  
Small Interfering Rnas ◽  
Viral Dna ◽  
Cyprinid Herpesvirus 3 ◽  
Interfering Rna ◽  
Fibroblastic Cells

Abstract A Cyprinid herpesvirus 3 infection of carp induces a disease which causes substantial losses in carp culture. Here we present the use of a possible strategy for the management of the virus infection RNA interference based on small interfering RNAs. As a result of in vitro studies, we found that a mixture of short interfering RNAs specific for viral DNA enzyme synthesis and capsid proteins of the CyHV-3 can be a potential inhibitor of virus replication in fibroblastic cells. This gives the basis for the development of a combinatorial RNA interference strategy to treat CyHV-3 infections.

scholarly journals The MAPK Kinase Kinase-1 Is Essential for Stress-Induced Pancreatic Islet Cell Death

Endocrinology ◽  
2008 ◽  
Vol 149 (6) ◽  
pp. 3046-3053 ◽  
Author(s):  
Dariush Mokhtari ◽  
Jason W. Myers ◽  
Nils Welsh
Keyword(s):  
Cell Death ◽  
Small Interfering Rna ◽  
Islet Cells ◽  
Β Cell ◽  
Pancreatic Islet Cell ◽  
Mapk Kinase ◽  
Transient Overexpression ◽  
Novel Therapeutic Strategies ◽  
Interfering Rna

The aim of the present investigation was to characterize the role of the MAPK kinase kinase-1 (MEKK-1) in stress-induced cell death of insulin producing cells. We observed that transient overexpression of the wild type MEKK-1 protein in the insulin-producing cell lines RIN-5AH and βTC-6 increased c-Jun N-terminal kinase (JNK) phosphorylation and augmented cell death induced by diethylenetriamine/nitroso-1-propylhydrazino)-1-propanamine (DETA/NO), streptozotocin (STZ), and hydrogen peroxide (H2O2). Furthermore, DETA/NO or STZ induced a rapid threonine phosphorylation of MEKK-1. Silencing of MEKK-1 gene expression in βTC-6 and human dispersed islet cells, using in vitro-generated diced small interfering RNA, resulted in protection from DETA/NO, STZ, H2O2, and tunicamycin induced cell death. Moreover, in DETA/NO-treated cells diced small interfering RNA-mediated down-regulation of MEKK-1 resulted in decreased activation of JNK but not p38 and ERK. Inhibition of JNK by treatment with SP600125 partially protected against DETA/NO- or STZ-induced cell death. In summary, our results support an essential role for MEKK-1 in JNK activation and stress-induced β-cell death. Increased understanding of the signaling pathways that augment or diminish β-cell MEKK-1 activity may aid in the generation of novel therapeutic strategies in the treatment of type 1 diabetes.

Effect of aquaporin 3 knockdown by RNA interference on antrum formation in sheep secondary follicles cultured in vitro

Zygote ◽  
2018 ◽  
Vol 26 (5) ◽  
pp. 350-358
Author(s):  
Marcela Pinheiro Paz ◽  
Francisca Geovania Canafístula de Sousa ◽  
Benner Geraldo Alves ◽  
Carlos Henrique Lobo ◽  
Antonia Débora Sales ◽  
...  
Keyword(s):  
Rna Interference ◽  
Basement Membrane ◽  
Granulosa Cell ◽  
Small Interfering Rna ◽  
Fluorescence Signal ◽  
Follicular Growth ◽  
Aquaporin 3 ◽  
Interfering Rna ◽  
Transfection Complex

SummaryThe objectives were to develop an effective protocol for transfection of ovine secondary follicles and to assess the effect of attenuating aquaporin 3 (AQP3) using a small interfering RNA (siRNA-AQP3) on antrum formation and follicular growth in vitro. Various combinations of Lipofectamine® volumes (0.5, 0.75 or 1.0 µl), fluorescent oligonucleotide (BLOCK-iT ™) concentrations (3.18, 27.12 or 36.16 nM) and exposure times (12, 14, 16, 18 or 20 h) were tested. The BLOCK-iT™ was replaced by siRNA-AQP3 in the transfection complex. Ovine secondary follicles were isolated and cultured in vitro for 6 days using standard protocols. Follicles were transfected on day 0 or 3 or on both days (0 and 3) and then cultured for an additional 3 or 6 days. As revealed by the fluorescence signal, the Lipofectamine®/BLOCK-iT™ complex (0.75 µl + 27.12 nM by 12 h of incubation) crossed the basement membrane and granulosa cell and reached the oocytes. In general, the rate of intact follicles was higher and the rate of antrum formation was lower in transfected follicles compared with control follicles. In conclusion, ovine secondary follicles can be successfully transfected during in vitro culture, and siRNA-mediated attenuation of AQP3 gene reduced antrum formation of secondary follicles.

scholarly journals Ketamine Activates Cell Cycle Signaling and Apoptosis in the Neonatal Rat Brain

2010 ◽  
Vol 112 (5) ◽  
pp. 1155-1163 ◽  
Author(s):  
Sulpicio G. Soriano ◽  
Qian Liu ◽  
Jing Li ◽  
Jia-Ren Liu ◽  
Xiao Hui Han ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Cyclin D1 ◽  
Small Interfering Rna ◽  
Caspase 3 ◽  
Primary Neurons ◽  
Cyclin Dependent Kinase ◽  
Cell Cycle Signaling ◽  
Interfering Rna

Background Prolonged exposure to ketamine results in accelerated neurodegeneration and neurocognitive deficits in the neonatal rats. Experimental models of neurodegeneration have implicated reentry of postmitotic neurons into the cell cycle, leading to cell death. The authors hypothesize that the ketamine-induced neuroapoptosis is partially due to aberrant cycle cell reentry. To explore this hypothesis, the authors characterized the effect of ketamine on the cell cycle signaling pathway in the developing rodent brain in vivo and in vitro. Methods Postnatal day 7 rat pups and primary neurons were used for the experiments. Each rat pup received five intraperitoneal doses of either saline or ketamine (5, 10, and 20 mg/kg/dose) at 90-min intervals over 6 h. Primary neurons were exposed to varying concentrations of ketamine to determine the dose and duration effects. The expression of cell cycle proteins (cyclin D1, cyclin-dependent kinase 4, and E2F1), Bcl2-interacting mediator of cell death (Bim), and activated caspase-3 was determined. The effect of cyclin D1 knockdown by small interfering RNA was also examined in primary neurons incubated in ketamine. Results Ketamine mediated a dose- and time-dependent increase in expression of cell cycle proteins and activated caspase-3. Cyclin D1, cyclin-dependent kinase 4, E2F1, Bim, and cleaved caspase-3 expression increased at 12 h and peaked at 24 h in vitro. Knockdown of cyclin D1 by small interfering RNA attenuated Bim and cleaved caspase-3 expression. Conclusion These findings support a model in which ketamine induces aberrant cell cycle reentry, leading to apoptotic cell death in the developing rat brain.

scholarly journals Cytotoxicity assessment, inflammatory properties, and cellular uptake of Neutraplex lipid-based nanoparticles in THP-1 monocyte-derived macrophages

2017 ◽  
Vol 4 ◽  
pp. 184954351774625 ◽  
Author(s):  
Eric Berger ◽  
Dalibor Breznan ◽  
Sandra Stals ◽  
Viraj J Jasinghe ◽  
David Gonçalves ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Small Interfering Rna ◽  
Human Monocyte ◽  
Antiretroviral Drugs ◽  
Small Interfering Rnas ◽  
Lactate Dehydrogenase Release ◽  
Drug Concentrations ◽  
Interfering Rna ◽  
Hiv 1

Current antiretroviral drugs used to prevent or treat human immunodeficiency virus type 1 (HIV-1) infection are not able to eliminate the virus within tissues or cells where HIV establishes reservoirs. Hence, there is an urgent need to develop targeted delivery systems to enhance drug concentrations in these viral sanctuary sites. Macrophages are key players in HIV infection and contribute significantly to the cellular reservoirs of HIV because the virus can survive for prolonged periods in these cells. In the present work, we investigated the potential of the lipid-based Neutraplex nanosystem to deliver anti-HIV therapeutics in human macrophages using the human monocyte/macrophage cell line THP-1. Neutraplex nanoparticles as well as cationic and anionic Neutraplex nanolipoplexes (Neutraplex/small interfering RNA) were prepared and characterized by dynamic light scattering. Neutraplex nanoparticles showed low cytotoxicity in CellTiter-Blue reduction and lactate dehydrogenase release assays and were not found to have pro-inflammatory effects. In addition, confocal studies showed that the Neutraplex nanoparticles and nanolipoplexes are rapidly internalized into THP-1 macrophages and that they can escape the late endosome/lysosome compartment allowing the delivery of small interfering RNAs in the cytoplasm. Furthermore, HIV replication was inhibited in the in vitro TZM-bl infectivity assay when small interfering RNAs targeting CXCR4 co-receptor was delivered by Neutraplex nanoparticles compared to a random small interfering RNA sequence. This study demonstrates that the Neutraplex nanosystem has potential for further development as a delivery strategy to efficiently and safely enhance the transport of therapeutic molecules into human monocyte-derived macrophages in the aim of targeting HIV-1 in this cellular reservoir.

scholarly journals A Small Interfering RNA Targeting Coxsackievirus B3 Protects Permissive HeLa Cells from Viral Challenge

2005 ◽  
Vol 79 (13) ◽  
pp. 8620-8624 ◽  
Author(s):  
Jeonghyun Ahn ◽  
Eun Seok Jun ◽  
Hui Sun Lee ◽  
Seung Yong Yoon ◽  
DongHou Kim ◽  
...  
Keyword(s):  
Cell Death ◽  
Viral Replication ◽  
Hela Cells ◽  
Small Interfering Rna ◽  
Antiviral Effect ◽  
Coxsackievirus B3 ◽  
Small Interfering Rnas ◽  
Antiviral Effects ◽  
Rna Targeting ◽  
Interfering Rna

ABSTRACT We examined the ability of small interfering RNAs (siRNAs) to disrupt infection by coxsackievirus B3 (CVB3). The incorporation of siRNAs dramatically decreased cell death in permissive HeLa cells in parallel with a reduction in viral replication. Three of four siRNAs had potent anti-CVB3 activity. The present study thus demonstrates that the antiviral effect is due to the downregulation of viral replication. In addition, an effective CVB3-specific siRNA had similar antiviral effects in other related enteroviruses possessing sequence homology in the targeted region. Because the CVB3-specific siRNA is effective against other enteroviruses, siRNAs have potential for a universal antienterovirus strategy.

scholarly journals Amphiphysin 1 Is Important for Actin Polymerization during Phagocytosis

2007 ◽  
Vol 18 (11) ◽  
pp. 4669-4680 ◽  
Author(s):  
Hiroshi Yamada ◽  
Emiko Ohashi ◽  
Tadashi Abe ◽  
Norihiro Kusumi ◽  
Shun-AI Li ◽  
...  
Keyword(s):  
Rna Interference ◽  
Sertoli Cells ◽  
Small Interfering Rna ◽  
Actin Polymerization ◽  
Actin Dynamics ◽  
Polymerization Process ◽  
Amphiphysin 1 ◽  
Phagocytic Cups ◽  
Interfering Rna ◽  
Constitutively Active

Amphiphysin 1 is involved in clathrin-mediated endocytosis. In this study, we demonstrate that amphiphysin 1 is essential for cellular phagocytosis and that it is critical for actin polymerization. Phagocytosis in Sertoli cells was induced by stimulating phosphatidylserine receptors. This stimulation led to the formation of actin-rich structures, including ruffles, phagocytic cups, and phagosomes, all of which showed an accumulation of amphiphysin 1. Knocking out amphiphysin 1 by RNA interference in the cells resulted in the reduction of ruffle formation, actin polymerization, and phagocytosis. Phagocytosis was also drastically decreased in amph 1 (−/−) Sertoli cells. In addition, phosphatidylinositol-4,5-bisphosphate–induced actin polymerization was decreased in the knockout testis cytosol. The addition of recombinant amphiphysin 1 to the cytosol restored the polymerization process. Ruffle formation in small interfering RNA-treated cells was recovered by the expression of constitutively active Rac1, suggesting that amphiphysin 1 functions upstream of the protein. These findings support that amphiphysin 1 is important in the regulation of actin dynamics and that it is required for phagocytosis.

scholarly journals Muscle-specific microRNA miR-206 promotes muscle differentiation

2006 ◽  
Vol 174 (5) ◽  
pp. 677-687 ◽  
Author(s):  
Hak Kyun Kim ◽  
Yong Sun Lee ◽  
Umasundari Sivaprasad ◽  
Ankit Malhotra ◽  
Anindya Dutta
Keyword(s):  
Antisense Oligonucleotide ◽  
Small Interfering Rna ◽  
Degradation Process ◽  
Microrna Target ◽  
C2c12 Myoblasts ◽  
Target Sites ◽  
The Many ◽  
Interfering Rna ◽  
In Vitro Transfection

Three muscle-specific microRNAs, miR-206, -1, and -133, are induced during differentiation of C2C12 myoblasts in vitro. Transfection of miR-206 promotes differentiation despite the presence of serum, whereas inhibition of the microRNA by antisense oligonucleotide inhibits cell cycle withdrawal and differentiation, which are normally induced by serum deprivation. Among the many mRNAs that are down-regulated by miR-206, the p180 subunit of DNA polymerase α and three other genes are shown to be direct targets. Down-regulation of the polymerase inhibits DNA synthesis, an important component of the differentiation program. The direct targets are decreased by mRNA cleavage that is dependent on predicted microRNA target sites. Unlike small interfering RNA–directed cleavage, however, the 5′ ends of the cleavage fragments are distributed and not confined to the target sites, suggesting involvement of exonucleases in the degradation process. In addition, inhibitors of myogenic transcription factors, Id1-3 and MyoR, are decreased upon miR-206 introduction, suggesting the presence of additional mechanisms by which microRNAs enforce the differentiation program.

Sign in / Sign up

Export Citation Format

Share Document