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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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Effect of blocking Rac1 expression in cholangiocarcinoma QBC939 cells BJMBR
Xudong,Liu; Guangyi,Wang.
Cholangiocarcinomas (CCs) are malignant tumors that originate from epithelial cells lining the biliary tree and gallbladder. Ras correlative C3 creotoxin substrate 1 (Rac1), a small guanosine triphosphatase, is a critical mediator of various aspects of endothelial cell functions. The objective of the present investigation was to study the effect of blocking Rac1 expression in CCs. Seventy-four extrahepatic CC (ECC) specimens and matched adjacent normal mucosa were obtained from the Department of Pathology, Inner Mongolia Medicine Hospital, between 2007 and 2009. Our results showed that the expression of Rac1 was significantly higher (53.12%) in tumor tissues than in normal tissues. Western blotting data indicated a significant reduction in Rac1-miRNA cell...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cholangiocarcinomas; RNAi; Rac1.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011000500015
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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 vivo RNA interference in oyster -vasa silencing inhibits germ cell development ArchiMer
Fabioux, Caroline; Corporeau, Charlotte; Quillien, Virgile; Favrel, Pascal; Huvet, Arnaud.
This study investigated the potential of RNA interference, which is technically challenging in bivalve mollusc species, to assess gene function in the oyster Crassostrea gigas. We designed dsRNA targeting the oyster vasa-like gene (Oyvlg), specifically expressed in oyster germ cells. In vivo injection of oyvl-dsRNA into the gonad provokes a knockdown phenotype corresponding to germ cell underproliferation and prematurely arrested meiosis througout the organ. The most severe phenotype observed is sterile. This knockdown phenotype is associated with a decrease in Oyvlg mRNA level of between 39% and 87%, and a strong reduction in OYVLG protein, to an undetectable level. Therefore, Oyvlg appears to be essential for germ cell development in Crassostrea gigas,...
Tipo: Text Palavras-chave: Vasa; RNAi; Marine bivalve; Germline; Crassostrea gigas.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6641.pdf
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Knockdown of E2f1 by RNA interference impairs proliferation of rat cells in vitro Genet. Mol. Biol.
Vasques,Luciana dos Reis; Pujiz,Regiane Simoni; Strauss,Bryan Eric; Krieger,José Eduardo.
E2F1 plays a key role in cell-cycle regulation in mammals, since its transcription factor activity controls genes required for DNA synthesis and apoptosis. E2F1 deregulation is a common feature among different tumor types and can be a major cause of cell proliferation. Thus, blocking E2F1 expression by RNA interference represents a promising therapeutic approach. In this study, the introduction of specific short hairpin RNAs (shRNAs) reduced E2f1 expression by up to 77%, and impaired rat glioma cell proliferation by approximately 70%, as compared to control cells. Furthermore, we investigated the expression of E2f1 target genes, Cyclin A and Cyclin E. Cyclin A was found to be down-regulated, whereas Cyclin E had similar expression to control cells,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; ShRNA; E2F1; Proliferation; Cancer.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000100005
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Molecular cloning, expression pattern, and putative cis-acting elements of a 4-coumarate:CoA ligase gene in bamboo (Neosinocalamus affinis) Electron. J. Biotechnol.
Cao,Ying; Hu,Shang-lian; Huang,Sheng-xiong; Ren,Peng; Lu,Xue-qin.
Background: 4-coumarate:CoA ligase (4CL) plays an important role at the divergence point from general phenylpropanoid metabolism to several branch pathways. Although 4CL sin higher plants have been extensively studied, little has known about the 4CL gene of bamboo. Results: In current study, a Na4CL gene putative encoding 4-coumarate:CoA ligase (4CL) and its 5’-flanking region were isolated from bamboo (Neosinocalamus affinis) by RACE-PCR and genomic DNA walker, respectively. Na4CL encodes a predicted protein of 557 amino acids, with conserved motifs of adenylate-forming enzymes. Phylogenetic analysis showed that Na4CL shared 62~85% identity with other known plant 4CLs, and cluster closely with some known 4CLs in monocots. Sequence analysis...
Tipo: Journal article Palavras-chave: Expression pattern; Neosinocalamus affinis; Promoter; RACE; RNAi.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500009
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Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs Electron. J. Biotechnol.
Stove,Veronique; Smits,Kaatje; Naessens,Evelien; Plum,Jean; Verhasselt,Bruno.
RNA interference (RNAi), mediated by short double-stranded RNAs, is a powerful mechanism for posttranscriptional gene silencing. Sustained expression of short hairpin RNA (shRNA) can be accomplished in mammalian cells by viral delivery systems. Using lentiviral constructs, stable gene silencing is established both in dividing and non-dividing cells. Targeting one single gene can lead to the development of escape mutants or may be insufficient to silence redundant pathways. Therefore, simultaneous targeting of multiple genes may be necessary. We have generated a lentiviral vector-based system for expression of multiple shRNAs from a single viral vector, which also encodes an EGFP reporter protein. We show that knock-down of each single gene from multiple...
Tipo: Journal article Palavras-chave: EGFP; Lentiviral gene transfer; Multiple knock-down; RNAi; Rho GTPases; ShRNA.
Ano: 2006 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013
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Multiple genes contribute to anhydrobiosis (tolerance to extreme desiccation) in the nematode Panagrolaimus superbus Genet. Mol. Biol.
Evangelista,Cláudia Carolina Silva; Guidelli,Giovanna Vieira; Borges,Gustavo; Araujo,Thais Fenz; Souza,Tiago Alves Jorge de; Neves,Ubiraci Pereira da Costa; Tunnacliffe,Alan; Pereira,Tiago Campos.
Abstract The molecular basis of anhydrobiosis, the state of suspended animation entered by some species during extreme desiccation, is still poorly understood despite a number of transcriptome and proteome studies. We therefore conducted functional screening by RNA interference (RNAi) for genes involved in anhydrobiosis in the holo-anhydrobiotic nematode Panagrolaimus superbus. A new method of survival analysis, based on staining, and proof-of-principle RNAi experiments confirmed a role for genes involved in oxidative stress tolerance, while a novel medium-scale RNAi workflow identified a further 40 anhydrobiosis-associated genes, including several involved in proteostasis, DNA repair and signal transduction pathways. This suggests that multiple genes...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Peroxiredoxin; Kinase; RNAi; Proteostasis.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572017000500790
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Non-specific activation of antiviral immunity and induction of RNA interference may engage the same pathway in the Pacific white leg shrimp Litopenaeus vannamei ArchiMer
Labreuche, Yannick; Veloso, Artur; De La Vega, Enrique; Gross, Paul S.; Chapman, Robert W.; Browdy, Craig L.; Warr, Gregory W..
Many questions remain unanswered regarding RNAi-based mechanisms and dsRNA-induced antiviral immune responses in penaeid shrimp. In this study, we report the characterization in the white leg shrimp Litopenaeus vannamei of RNAi pathway associated proteins Lv-Ago 1 and Lv-Ago 2, two members of the Argonaute family of proteins, as well as Lv-sid 1, the first shrimp homologue of Sid-1, a membrane channel-forming protein implicated in the cellular import of dsRNA. To decipher their functional implication in RNAi-related phenomena, we monitored their relative expression following stimulation by specific and non-specific RNA duplexes of diverse length. The findings show that the length of small RNA duplexes plays a critical role in the activation of both...
Tipo: Text Palavras-chave: Shrimp; Litopenaeus vannamei; RNAi; DsRNA; Argonaute; Sid-1; Antiviral immunity.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00015/12652/9587.pdf
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Sheep 7SK promoter for short hairpin RNA expression Electron. J. Biotechnol.
Ni,Wei; Hu,Shengwei; Hazi,Wureli; Wang,Yuanzhi; Qiao,Jun; Yan,Ren; Chen,Chuangfu.
Background: Gene silencing mediated by small interfering RNA (siRNA) has become a powerful biological tool for the regulation of gene expression. For the synthesis of siRNA by vector-based expression systems, several mammalian small nuclear RNA (snRNA) promoters have been cloned and shown different transcriptional efficiencies. Results: In this study, we identified a sheep 7SK snRNA (s7SK) promoter based on the highly conserved polymerase III promoter elements. Promoter activity was measured by promoter-driven shRNA expression to suppress expression of an exogenous reporter gene and endogenous sheep gene. Conclusions: The knock down assay demonstrated that the s7SK induced more stronger inhibition effect than human U6 and H1 promoters. The use of this...
Tipo: Journal article Palavras-chave: 7SK promoter; RNAi; Sheep; ShRNA.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000200010
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Silencing HIF-1α reduces the adhesion and secretion functions of acute leukemia hBMSCs BJMBR
Dong-Feng,Zeng; Ting,Liu; Cheng,Chang; Xi,Zhang; Xue,Liang; Xing-Hua,Chen; Pei-Yan,Kong.
Hypoxia inducible factor-1α (HIF-1α) is an important transcription factor, which plays a critical role in the formation of solid tumor and its microenviroment. The objective of the present study was to evaluate the expression and function of HIF-1α in human leukemia bone marrow stromal cells (BMSCs) and to identify the downstream targets of HIF-1α. HIF-1α expression was detected at both the RNA and protein levels using real-time PCR and immunohistochemistry, respectively. Vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1α (SDF-1α) were detected in stromal cells by enzyme-linked immunosorbent assay. HIF-1α was blocked by constructing the lentiviral RNAi vector system and infecting the BMSCs. The Jurkat cell/BMSC co-cultured system...
Tipo: Info:eu-repo/semantics/article Palavras-chave: HIF-1α; Human bone marrow stromal cells (hBMSCs); Leukemia; RNAi; SDF-1α; VEGF.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2012001000004
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Stable silencing of β-lactoglobulin (BLG) gene by lentivirus-mediated RNAi in goat fetal fibroblasts Genet. Mol. Biol.
Zhang,Shumin; Xiong,Kai; Xie,Zhourui; Nan,Wenting; Liu,Honglin; Chen,Jie.
β-lactoglobulin (BLG), a dominant allergen in goat milk, is difficult to remove by traditional biochemical methods. Its elimination from goat milk by genetic modification therefore poses a major challenge for modern goat breeders. A shRNA targeting BLG mRNA with high interference efficiency was identified, with which lentiviral vectors were used for mediating stable shRNA interference in goat-fetal fibroblast cells. Apart from high efficiency in the knockdown of BLG expression in these cells, lentivector-mediated RNAi manifested stable integration into the goat genome itself. Consequently, an in vitro model for goat BLG-content control was compiled, and a goat-cell line for accompanying transgenetic goat production created.
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; ShRNA; Lentivirus; BLG; Goat.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000400020
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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 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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