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Registros recuperados: 10
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A practical approach to the understanding and teaching of RNA silencing in plants Electron. J. Biotechnol.
Bazzini,Ariel A; Mongelli,Vanesa C; Hopp,H. Esteban; del Vas,Mariana; Asurmendi,Sebastián.
Gene silencing, also called RNA interference (RNAi) is a specific mechanism of RNA degradation involved in gene regulation, development and defense in eukaryotic organisms. It became an important subject in the teaching programs of molecular biology, genetics and biotechnology courses in the last years. The aim of this work is to provide simple and inexpensive assays to understand and teach gene silencing using plants as model systems. The use of transient and permanent transgenic plants for expressing reporter genes, like those derived from jellyfish green fluorescent protein (gfp) encoding gene, provides a nice, colorful and conclusive image of gene silencing. Three experimental approaches to evidence RNA silencing are depicted. In the first approach...
Tipo: Journal article Palavras-chave: Gene silencing; GFP; HC-Pro; PDS; Protocols; PTGS; Teaching; Transgenic; Transient expression; TRV; VIGS.
Ano: 2007 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200002
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Evaluation of the uniformity and stability of T-DNA integration and gene expression in transgenic apple plants Electron. J. Biotechnol.
Flachowsky,Henryk; Riedel,Marko; Reim,Stefanie; Hanke,Magda-Viola.
The generation of transgenic apple plants relies on the molecular analysis of transgene integration and expression based on polymerase chain reaction (PCR) analysis, blotting techniques and enzymatic assays on vitro leaves of putative transgenic regenerates. In order to assess the uniformity and the stability of transfer DNA (T-DNA) integration and gene expression, we studied 26 transgenic apple lines carrying the attacin E gene from Hyalophora cecropia, the β-glucuronidase gene, and the nptII gene. Plants were evaluated using standard molecular techniques, such as PCR, Southern blot, reverse transcription PCR (RT-PCR) and Enzyme Linked Immunosorbent Assay (ELISA), and propagated in vitro on non-selective antibiotic-free media for four years to...
Tipo: Journal article Palavras-chave: Agrobacterium tumefaciens; Apple; Chimeric tissue; Gene silencing; Malus domestica; T-DNA leakage.
Ano: 2008 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100003
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Genetic machinery for RNA silencing and defense against viruses in Citrus Genet. Mol. Biol.
Benedito,Vagner Augusto; Faria,Laura; Freitas-Astúa,Juliana; Figueira,Antonio.
RNA silencing mechanisms are conserved throughout eukaryotic evolution, possibly due to their importance in viral resistance and other aspects of cell biology. Here, we explored the Citrus EST (CitEST) database in search of sequences related to the most important known genes involved in RNA silencing. Transcripts strongly matching Argonaute (AGO), Dicer-like (DCL), Hua enhancer (HEN), and RNA-dependent RNA Polymerase (RdRP) were found in many of the citrus libraries. The reads were clustered and quantified. This shows that post-transcriptional gene silencing apparatus is active in citrus. It seems plausible that a better understanding of the players of RNA silencing in Citrus spp. and related genera may help create new tools to defeat the viral diseases...
Tipo: Info:eu-repo/semantics/article Palavras-chave: CitEST; Gene silencing; PTGS.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000500027
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In Silico Identification of MicroRNAs with B/CYDV Gene Silencing Potential BABT
Bouallègue,Maryem; Bouktila,Dhia; Mezghani-Khemakhem,Maha; Capy,Pierre; Makni,Mohamed.
ABSTRACT Computational investigation of a set of publicly available plant microRNAs revealed 19 barley- and other plants-encoded miRNAs and their near-complement reverse sequences (miRNA*) that have potential to bind all B/CYDV open reading frames (ORFs) except ORF0 and ORF6. These miRNAs/miRNAs*, their binding positions and targets are discussed in the context of biological protection of cereals against B/CYDV, based on antiviral silencing.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Barley/Cereal Yellow Dwarf virus (B/CYDV); MicroRNA; Gene silencing; Host defense.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100437
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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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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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Targeting PPM1D by lentivirus-mediated RNA interference inhibits the tumorigenicity of bladder cancer cells BJMBR
Wang,W.; Zhu,H.; Zhang,H.; Zhang,L.; Ding,Q.; Jiang,H..
Protein phosphatase magnesium/manganese-dependent 1D (PPM1D) is a p53-induced phosphatase that functions as a negative regulator of stress response pathways and has oncogenic properties. However, the functional role ofPPM1D in bladder cancer (BC) remains largely unknown. In the present study, lentivirus vectors carrying small hairpin RNA (shRNA) targeting PPM1D were used to explore the effects ofPPM1D knockdown on BC cell proliferation and tumorigenesis. shRNA-mediated knockdown of PPM1D significantly inhibited cell growth and colony forming ability in the BC cell lines 5637 and T24. Flow cytometric analysis showed that PPM1D silencing increased the proportion of cells in the G0/G1 phase. Downregulation of PPM1Dalso inhibited 5637 cell tumorigenicity in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Protein phosphatase magnesium/manganese-dependent 1D; Bladder cancer; Gene silencing; RNA interference; Proliferation.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2014001201044
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The post-transcriptional gene silencing pathway in Eucalyptus Genet. Mol. Biol.
Sassaki,Flávio Tetsuo; Campos-Pereira,Tiago; Maia,Ivan de Godoy.
Post-transcriptional gene silencing (PTGS) is a conserved surveillance mechanism that identifies and cleaves double-stranded RNA molecules and their cellular cognate transcripts. The RNA silencing response is actually used as a powerful technique (named RNA interference) for potent and specific inhibition of gene expression in several organisms. To identify gene products in Eucalyptus sharing similarities with enzymes involved in the PTGS pathway, we queried the expressed sequence tag database of the Brazilian Eucalyptus Genome Sequence Project Consortium (FORESTs) with the amino acid sequences of known PTGS-related proteins. Among twenty-six prospected genes, our search detected fifteen assembled sequences encoding products presenting high level of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Gene silencing; PTGS; RNA silencing; EST; Eucalyptus.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572005000400003
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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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Traditional and transgenic strategies for controlling tomato-infecting begomoviruses Trop. Plant Pathol.
FREITAS-ASTÚA,JULIANA; PURCIFULL,DAN E.; POLSTON,JANE E.; HIEBERT,ERNEST.
Viruses of to the family Geminiviridae are considered some of the most important pathogens in tropical and subtropical regions of the world. Members of one Geminiviridae genus, Begomovirus, have been causing severe losses, particularly in tomato (Lycopersicon esculentum) production in the Americas and the Caribbean. Several new begomoviruses have been reported in the region and, at least one, Tomato yellow leaf curl virus (TYLCV), has been brought in from the Old World via infected transplants. In addition, the recombination events that are playing an important role in Begomovirus diversity have increased the complexity of their control. This scenario has led to the search for control measures that go beyond traditional host genetic resistance, chemical...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Whitefly-transmitted geminivirus; Gene silencing; Plant transformation; Tomato begomovirus; Geminivirus control; Geminiviridae.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582002000500001
Registros recuperados: 10
Primeira ... 1 ... Última
 

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