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Registros recuperados: 15
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Analysis of energetically biased transcripts of viruses and transposable elements Genet. Mol. Biol.
Secolin,Rodrigo; Pascoal,Vinícius D'Ávila Bitencourt; Lopes-Cendes,Iscia; Pereira,Tiago Campos.
RNA interference (RNAi) is a natural endogenous process by which double-stranded RNA molecules trigger potent and specific gene silencing in eukaryotic cells and is characterized by target RNA cleavage. In mammals, small interfering RNAs (siRNAs) are the trigger molecules of choice and constitute a new class of RNA-based antiviral agents. In an efficient RNAi response, the antisense strand of siRNAs must enter the RNA-induced silencing complex (RISC) in a process mediated by thermodynamic features. In this report, we hypothesize that silent mutations capable of inverting thermodynamic properties can promote resistance to siRNAs. Extensive computational analyses were used to assess whether continuous selective pressure that promotes such mutations could...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; SiRNA; Synonymous mutation; Viral evolution.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000500022
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Effects of c-FLIPL Knockdown in Cervical Uterine Carcinoma Cell Lines International Journal of Morphology
Ili,Carmen G; Brebi,Priscilla; Garcia,Patricia; Leal,Pamela; Lopez,Jaime; Tapia,Oscar; Letelier,Pablo; Weber,Helga; Castillo,Jonathan; Roa,Juan C.
Overexpression of Short and Raji variants of Cellular FLICE-like inhibitory protein (c-FLIP) is capable of inhibiting apoptosis, while the function of the Long isoform depends of c-FLIPL concentration in cells. The aim of this study was to determine the effects of c-FLIPL knockdown in cervical cell lines. SiHa, C-4I and C-33A cervical cancer cell lines were analyzed. c-FLIPL level expression was determined by quantitative real-time PCR and western blotting. c-FLIPL was transiently downregulated by siRNA. The effects of knockdown of c-FLIPL on cell viability, proliferation and apoptosis were assessed by comparing with scrambled siRNA-transfected cells. SiHa and C-4I c-FLIPL knockdown cells showed increased viability compared with scrambled siRNA-transfected...
Tipo: Journal article Palavras-chave: Apoptosis; Proliferation; Cervical cancer; SiRNA; C-FLIPL.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022015000200036
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Effects of PSMA1 on the differentiation and lipid deposition of bovine preadipocytes R. Bras. Zootec.
Li,Sheng; Chen,Chengzhen; Jiang,Hao; Zhang,Jiabao; Zhou,Qian; Gao,Yan; Yuan,Bao; Zhang,Mingjun.
ABSTRACT In this study, our goal was to clarify the role of proteasomal subunit α-1 (PSMA1) in both the differentiation of preadipocytes and the accumulation of lipids in adipocytes. Preadipocytes from healthy one-day-old calves were collected, isolated, and cultured in vitro. The expression pattern of the PSMA1 gene was explored during the differentiation of bovine preadipocytes firstly. Then, the expression of the PSMA1 gene was inhibited by transfection of a chemically synthesized small interfering RNA (siRNA) before differentiation. After induction of differentiation, the mRNA levels of key regulating genes involved in preadipocyte differentiation and the lipid content of mature adipocytes with and without inhibition of PSMA1 were detected by qRT-PCR...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Gene expression; SiRNA; Transfection; Triglyceride.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982019000100309
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Evaluation of short-interfering RNAs treatment in experimental rabies due to wild-type virus BJID
Appolinario,Camila Michele; Allendorf,Susan Dora; Peres,Marina Gea; Fonseca,Clovis Reynaldo; Vicente,Acacia Ferreira; Antunes,João Marcelo Azevedo de Paula; Pantoja,José Carlos Figueiredo; Megid,Jane.
ABSTRACTWe have evaluated the efficacy of short-interfering RNAs targeting the nucleoprotein gene and also the brain immune response in treated and non-treated infected mice. Mice were inoculated with wild-type virus, classified as dog (hv2) or vampire bat (hv3) variants and both groups were treated or leaved as controls. No difference was observed in the lethality rate between treated and non-treated groups, although clinical evaluation of hv2 infected mice showed differences in the severity of clinical disease (p = 0.0006). Evaluation of brain immune response 5 days post-inoculation in treated hv2 group showed no difference among the analyzed genes, whereas after 10 days post-inoculation there was increased expression of 2',5'-oligoadenylate synthetase...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Rabies; Dog virus; Bat virus; SiRNA.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000500453
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Gene silencing: concepts, applications, and perspectives in woody plants Scientia Agricola
Souza,Amancio José de; Mendes,Beatriz Madalena Januzzi; Mourão Filho,Francisco de Assis Alves.
RNA interference, transcriptional gene silencing, virus induced gene silencing, and micro RNAs comprise a series of mechanisms capable of suppressing gene expression in plants. These mechanisms reveal similar biochemical pathways and appear to be related in several levels. The ability to manipulate gene silencing has produced transgenic plants able to switch off endogenous genes and invading nucleic acids. This powerful biotechnological tool has provided plant breeders and researchers with great opportunity to accelerate breeding programs and developmental studies in woody plants. This research work reports on gene silencing in woody plants, and discuss applications and future perspectives.
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; MiRNA; SiRNA; Genetic transformation; Virus resistance.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162007000600014
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In vitro modulation of Bcl-2 levels in small cell lung cancer cells: effects on cell viability BJMBR
Santos,A.O.; Pereira,J.P.; Pedroso de Lima,M.C.; Simões,S.; Moreira,J.N..
Small cell lung cancer (SCLC) is an aggressive disease, representing 15% of all cases of lung cancer, has high metastatic potential and low prognosis that urgently demands the development of novel therapeutic approaches. One of the proposed approaches has been the down-regulation of BCL2, with poorly clarified and controversial therapeutic value regarding SCLC. The use of anti-BCL2 small interfering RNA (siRNA) in SCLC has never been reported. The aim of the present study was to select and test the in vitro efficacy of anti-BCL2 siRNA sequences against the protein and mRNA levels of SCLC cells, and their effects on cytotoxicity and chemosensitization. Two anti-BCL2 siRNAs and the anti-BCL2 G3139 oligodeoxynucleotide (ODN) were evaluated in SCLC cells by...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Small cell lung cancer; Gene silencing; Bcl-2; SiRNA; Antisense ODN; Flow cytometry; Chemosensitization.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010001000013
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Inhibition of Rho-associated kinases disturbs the collective cell migration of stratified TE-10 cells Biol. Res.
Mikami,Taro; Yoshida,Keiichiro; Sawada,Hajime; Esaki,Michiyo; Yasumura,Kazunori; Ono,Michio.
BACKGROUND: The collective cell migration of stratified epithelial cells is considered to be an important phenomenon in wound healing, development, and cancer invasion; however, little is known about the mechanisms involved. Furthermore, whereas Rho family proteins, including RhoA, play important roles in cell migration, the exact role of Rho-associated coiled coil-containing protein kinases (ROCKs) in cell migration is controversial and might be cell-type dependent. Here, we report the development of a novel modified scratch assay that was used to observe the collective cell migration of stratified TE-10 cells derived from a human esophageal cancer specimen. RESULTS: Desmosomes were found between the TE-10 cells and microvilli of the surface of the cell...
Tipo: Journal article Palavras-chave: Collective cell migration; Stratified epithelium; TE-10 cells; Rho-associated coiled coil-containing protein kinase; SiRNA.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100048
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Integrin-β1, not integrin-β5, mediates osteoblastic differentiation and ECM formation promoted by mechanical tensile strain Biol. Res.
Zeng,Qiangcheng; Guo,Yong; Liu,Yongming; Li,Ruixin; Zhang,Xinchang; Liu,Lu; Wang,Yang; Zhang,Xizheng; Zou,Xianqiong.
BACKGROUND: Mechanical strain plays a great role in growth and differentiation of osteoblast. A previous study indicated that integrin-β (β1, β5) mediated osteoblast proliferation promoted by mechanical tensile strain. However, the involvement of integrin-β in osteoblastic differentiation and extracellular matrix (ECM) formation induced by mechanical tensile strain, remains unclear. RESULTS: After transfection with integrin-β1 siRNA or integrin-β5 siRNA, mouse MC3T3-E1 preosteoblasts were cultured in cell culture dishes and stimulated with mechanical tensile strain of 2500 microstrain (µε) at 0.5 Hz applied once a day for 1 h over 3 or 5 consecutive days. The cyclic tensile strain promoted...
Tipo: Journal article Palavras-chave: Tensile strain; Osteoblast; Extracellular matrix; Integrin; SiRNA.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100025
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Metatranscriptomic analysis of small RNAs present in soybean deep sequencing libraries Genet. Mol. Biol.
Molina,Lorrayne Gomes; Cordenonsi da Fonseca,Guilherme; Morais,Guilherme Loss de; de Oliveira,Luiz Felipe Valter; Carvalho,Joseane Biso de; Kulcheski,Franceli Rodrigues; Margis,Rogerio.
A large number of small RNAs unrelated to the soybean genome were identified after deep sequencing of soybean small RNA libraries. A metatranscriptomic analysis was carried out to identify the origin of these sequences. Comparative analyses of small interference RNAs (siRNAs) present in samples collected in open areas corresponding to soybean field plantations and samples from soybean cultivated in greenhouses under a controlled environment were made. Different pathogenic, symbiotic and free-living organisms were identified from samples of both growth systems. They included viruses, bacteria and different groups of fungi. This approach can be useful not only to identify potentially unknown pathogens and pests, but also to understand the relations that...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Next generation sequencing; Small RNA; SiRNA; Molecular markers.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200010
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RNA interference inhibits herpes simplex virus type 1 isolated from saliva samples and mucocutaneous lesions BJID
Silva,Amanda Perse da; Lopes,Juliana Freitas; Paula,Vanessa Salete de.
The aim of this study was to evaluate the use of RNA interference to inhibit herpes simplex virus type-1 replication in vitro. For herpes simplex virus type-1 gene silencing, three different small interfering RNAs (siRNAs) targeting the herpes simplex virus type-1 UL39 gene (sequence si-UL 39-1, si-UL 39-2, and si-UL 39-3) were used, which encode the large subunit of ribonucleotide reductase, an essential enzyme for DNA synthesis. Herpes simplex virus type-1 was isolated from saliva samples and mucocutaneous lesions from infected patients. All mucocutaneous lesions' samples were positive for herpes simplex virus type-1 by real-time PCR and by virus isolation; all herpes simplex virus type-1 from saliva samples were positive by real-time PCR and 50% were...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Herpes simplex virus type 1; Real-time PCR; SiRNA.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702014000400441
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Short-interfering RNAs as antivirals against rabies BJID
Brandão,Paulo Eduardo; Castilho,Juliana Galera; Fahl,Willian; Carnieli Jr.,Pedro; Oliveira,Rafael de Novaes; Macedo,Carla Isabel; Carrieri,Maria Luiza; Kotait,Ivanete.
This study aimed to test in vitro a RNA-interference based antiviral approach for rabies with short-interfering RNAs (siRNAs) against rabies virus nucleoprotein mRNA. BHK-21 cells were infected with serial dilutions of PV rabies virus strain and transfected with a pool of three siRNAs. Direct immunofluorescence staining showed a 5-time decrease in virus titer when compared to a non-treated plate, showing a promising new approach to the development of antivirals for rabies treatment.
Tipo: Info:eu-repo/semantics/article Palavras-chave: SiRNA; Rabies; Antiviral.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702007000200011
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SSD - a free software for designing multimeric mono-, bi- and trivalent shRNAs Genet. Mol. Biol.
Carli,Gabriel José de; Rotela,Abdon Troche; Lubini,Greice; Contiliani,Danyel Fernandes; Candia,Nidia Benítez; Depintor,Thiago S.; Abreu,Fabiano Carlos Pinto de; Simões,Zilá Luz Paulino; Ríos,Danilo Fernández; Pereira,Tiago Campos.
Abstract RNA interference (RNAi) is a powerful gene silencing technology, widely used in analyses of reverse genetics, development of therapeutic strategies and generation of biotechnological products. Here we present a free software tool for the rational design of RNAi effectors, named siRNA and shRNA designer (SSD). SSD incorporates our previously developed software Strand Analysis to construct template DNAs amenable for the large scale production of mono-, bi- and trivalent multimeric shRNAs, via in vitro rolling circle transcription. We tested SSD by creating a trivalent multimeric shRNA against the vitellogenin gene of Apis mellifera. RT-qPCR analysis revealed that our molecule promoted a decrease in more than 50% of the target mRNA, in a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Gene silencing; SiRNA; Multimeric; ShRNA; Free software.
Ano: 2020 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572020000100802
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Strand Analysis, a free online program for the computational identification of the best RNA interference (RNAi) targets based on Gibbs free energy Genet. Mol. Biol.
Pereira,Tiago Campos; Bittencourt,Vinícius D'Ávila Pascoal; Secolin,Rodrigo; Rocha,Cristiane de Souza; Maia,Ivan de Godoy; Lopes-Cendes,Iscia.
The RNA interference (RNAi) technique is a recent technology that uses double-stranded RNA molecules to promote potent and specific gene silencing. The application of this technique to molecular biology has increased considerably, from gene function identification to disease treatment. However, not all small interfering RNAs (siRNAs) are equally efficient, making target selection an essential procedure. Here we present Strand Analysis (SA), a free online software tool able to identify and classify the best RNAi targets based on Gibbs free energy (deltaG). Furthermore, particular features of the software, such as the free energy landscape and deltaG gradient, may be used to shed light on RNA-induced silencing complex (RISC) activity and RNAi mechanisms,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; SiRNA; SiRNA design; Software.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000600030
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The effect of down-regulation of Smad3 by RNAi on hepatic stellate cells and a carbon tetrachloride-induced rat model of hepatic fibrosis BJMBR
Wang,Z.R.; Wang,J.H.; Hu,C.L.; Cao,W.G.; Shen,X.J.; Wu,M.Y.; Shen,L.; Wu,S.L..
Searching for effective Smad3 gene-based gene therapies for hepatic fibrosis, we constructed siRNA expression plasmids targeting the rat Smad3 gene and then delivered these plasmids into hepatic stellate cells (HSCs). The effect of siRNAs on the mRNA levels of Smad2, Smad3, Smad4, and collagens I-α1, III-α1 and IV-α1 (Colα1, Col3α1, Col4α1, respectively) was determined by RT-PCR. Eighty adult male Sprague-Dawley rats were randomly divided into three groups. Twice a week for 8 weeks, the untreated hepatic fibrosis model (N = 30) and the treated group (N = 20) were injected subcutaneously with 40% (v/v) carbon tetrachloride (CCl4)-olive oil (3 mL/kg), and the normal control group (N = 30) was injected with olive oil (3 mL/kg). In the 4th week, the treated...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Smad3; SiRNA; Hepatic fibrosis; Carbon tetrachloride.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011000200002
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The RNA interference revolution BJMBR
Lenz,G..
The discovery of double-stranded RNA-mediated gene silencing has rapidly led to its use as a method of choice for blocking a gene, and has turned it into one of the most discussed topics in cell biology. Although still in its infancy, the field of RNA interference has already produced a vast array of results, mainly in Caenorhabditis elegans, but recently also in mammalian systems. Micro-RNAs are short hairpins of RNA capable of blocking translation, which are transcribed from genomic DNA and are implicated in several aspects from development to cell signaling. The present review discusses the main methods used for gene silencing in cell culture and animal models, including the selection of target sequences, delivery methods and strategies for a successful...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; SiRNA; MiRNA; Gene silencing; Knockdown.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005001200003
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